US Pat. No. 9,353,311

COMPOSITE VISIBLE COLORANT AND METHOD FOR QUANTITATIVE AMPLIFICATION

Bio-Rad Laboratories, Inc...

1. A composite colorant formulation comprising:
a visually-detectable dye; and
an internal reference molecule comprising a first fluorophore, which first fluorophore upon excitation emits energy at a specific
wavelength range;

wherein:
the visually-detectable dye partially absorbs energy at the specific wavelength range; and
the formulation does not comprise sufficient components for target nucleic acid amplification if added to a target nucleic
acid and primers.

US Pat. No. 9,403,294

METHOD OF MAKING A DROPLET-GENERATING DEVICE

Bio-Rad Laboratories, Inc...

1. A method of making a droplet-generating device, the method comprising:
producing a first droplet-generating device including a molded portion created at least in part with a mold and also including
a plurality of droplet generators each formed at least in part by the molded portion;

generating a set of droplets with each of one or more of the droplet generators;
determining a property of at least one set of generated droplets;
modifying the mold based on the property; and
producing a second droplet-generating device including a molded portion created at least in part with the modified mold;
wherein the mold defines a projecting portion including a plurality of intersecting ridges that create a plurality of intersecting
grooves in the molded portion, and wherein the step of modifying the mold includes a step of removing material from the projecting
portion of the mold.

US Pat. No. 9,354,179

OPTICAL DETECTION SYSTEM FOR LIQUID SAMPLES

Bio-Rad Laboratories Inc....

1. A tip for use in an optical detection system to analyze an analyte in a fluid sample drawn into the tip, using light reflected
from a detection surface inside the tip that the analyte binds to, comprising a first detection surface and a second detection
surface located in a same flow path with no controllable valve separating them, wherein the first and second detection surfaces
have different surface chemistries, and wherein the second detection surface has a surface chemistry that blocks it from binding
to a ligand that binds to the analyte, while the first detection surface has a surface chemistry that allows it to bind to
the ligand, wherein said surface chemistry of said first detection surface comprises a capturing agent selected from the group
consisting of avidin, an avidin derivative, protein A, protein G, a secondary antibody and Ni(II) activated complexes of nitrilo
triacetic acid, or comprises a functional group comprising one or more of active carboxylic ester, epoxide, aldehyde, and
acrylate , hydroxyl, alkyl ester, carboxylic acid, sulfate, sulfonate and phosphonate, such that said analyte binds to said
detection surface by binding to a ligand that binds to said surface chemistry, and
wherein said first detection surface has said capturing agent, and wherein said second detection surface, does not comprise
a capturing agent for affinity-based binding of the ligand.

US Pat. No. 9,234,875

SIMULTANEOUS PURIFICATION OF CELL COMPONENTS

BIO-RAD LABORATORIES, INC...

1. A method of purifying at least one cellular component from a biological sample comprising one or more cells or a lysate
thereof, wherein the one or more cells or lysate thereof comprises a plurality of cellular components of the one or more cells,
the method comprising,
providing into a chamber the sample; and
generating a pH gradient or pH step in the chamber with one or more proton injector(s) and/or hydroxide injector(s), such
that at least two different cellular components from the plurality of cellular components are positioned in different positions
in the chamber based on the isoelectric point (pI) of the at least two different components, thereby purifying at least one
cellular component from a different cellular component,

wherein the plurality of cellular components are electrophoretically moved through the inside of the chamber.
US Pat. No. 9,422,602

METHODS AND COMPOSITIONS FOR DETERMINING NUCLEIC ACID DEGRADATION

Bio-Rad Laboratories, Inc...

1. A method of detecting degradation of mRNA, the method comprising:
(a) contacting a sample comprising mRNA with at least one agent to covalently attach an oligonucleotide label to a first end
of RNA molecules having a first end structure and to covalently attach an oligonucleotide label to a second end of mRNA molecules
having a second end structure;

(b) separating the sample into a plurality of spatially isolated partitions;
(c) enumerating a first number of spatially isolated partitions comprising an oligonucleotide label covalently attached to
the first end of an mRNA molecule and an oligonucleotide label covalently attached to the second end of an mRNA molecule,
and a second number of spatially isolated partitions comprising an oligonucleotide label covalently attached only to the first
end or only to the second end of an mRNA molecule; and

(d) determining an amount of degradation for the mRNA based on the enumerating, wherein the step of determining comprises
comparing the first number and the second number to one another.

US Pat. No. 9,321,012

ELECTRONIC PROTEIN FRACTIONATION

Bio-Rad Laboratories, Inc...

1. An apparatus, comprising
a chamber divided into a first sub-chamber and a second sub-chamber by a dividing membrane, wherein the dividing membrane
blocks or substantially blocks flow of fluid between the first and second sub-chamber; and wherein

the first sub-chamber is in electrical and fluid communication with a first ion injector extractor comprising a first electrode;
the second sub-chamber is in electrical and fluid communication with a second ion injector extractor comprising a second electrode;
and
the second sub-chamber comprises an outlet,
wherein the ion injector/extractors each comprise:
a. a compartment in fluid communication with a sub-chamber and divided from the sub-chamber by an anion selective membrane,
wherein the anion selective membrane is permeable to small anions but not larger molecules; and/or

b. a compartment in fluid communication with the sub-chamber and divided from the sub-chamber by a bipolar membrane, wherein
the bipolar membrane is permeable to small ions but not larger molecules; and/or

c. compartment in fluid communication with a sub-chamber and divided from the sub-chamber by a cation selective membrane,
wherein the cation selective membrane is permeable to small cations but not larger molecules.

US Pat. No. 9,217,174

MAGNETIC LYSIS METHOD AND DEVICE

Akonni Biosystems, Inc., ...

1. A method for lysing cells within a vessel, said method comprising:
placing said vessel on a surface, wherein said vessel comprises one or more cells suspended in a liquid medium, one or more
magnetic stirrers, and a plurality of beads; and

rotating a cylinder shaped magnet along an axis, wherein said axis passes the center of said cylinder shaped magnet, is in
the proximity of said vessel and is parallel to the surface said vessel resides on so that the vessel is within operational
range of a rotating magnetic field produced by said cylinder shaped magnet, and wherein said rotating said cylinder shaped
magnet causes said one or more magnetic stirrers in said vessel to collide with said beads with enough force to cause disintegration
of said one or more cells, thereby lysing said one or more cells within said vessel.

US Pat. No. 9,427,737

METHODS AND COMPOSITIONS FOR USING OILS FOR ANALYSIS AND DETECTION OF MOLECULES

Bio-Rad Laboratories, Inc...

1. A composition comprising:
aqueous droplets; and
a homogeneous oil mixture forming a continuous phase that encapsulates each of the aqueous droplets, the oil mixture including
(a) a silicone oil and (b) a fluorinated oil;

wherein the concentration of said silicone oil in the oil mixture is at least about 50% by weight, and
wherein the aqueous droplets have a neutral buoyancy in the oil mixture.

US Pat. No. 9,322,830

METHOD FOR STUDYING TRANSPORT OF AN AGENT ACROSS A BILAYER MEMBRANE IN BIOANALYTICAL SENSOR APPLICATIONS

Bio-Rad Laboratories, Inc...

1. A method comprising:
a. providing at least one surface with a bilayer structure tethered to the at least one surface, said bilayer structure enclosing
a detection volume,

b. contacting the bilayer structure with at least one agent to be analysed,
c. detecting a change in refractive index only in the detection volume enclosed by the bilayer structure resulting from transportation
of the at least one agent across a membrane of the bilayer structure, wherein the change in refractive index is detected with
at least one sensor selected from the group consisting of a surface plasmon resonance sensor, an ellipsometry sensor, and
an optical waveguide laser spectroscopy sensor, and

d. determining a rate at which the at least one agent is released from the bilayer structure using the detected change in
the refractive index of the detection volume.

US Pat. No. 9,216,392

SYSTEM FOR FORMING AN ARRAY OF EMULSIONS

Bio-Rad Laboratories, Inc...

1. A system for forming an array of emulsions, comprising:
a plate including an array of emulsion production units, each unit including
at least one first input well to hold a continuous phase for an emulsion,
a second input well to hold a dispersed phase for an emulsion, and
an output well connected to the first and second input wells by a set of channels that form a channel junction, the set of
channels including at least two input channels extending separately from the input wells to the channel junction and an output
channel extending from the channel junction to the output well, each channel of the set of channels being circumferentially
bounded; and

a vacuum or pressure source configured to be connected operatively to wells of the plate to form a pressure drop between the
input wells and the output well of each unit to drive the continuous phase and the dispersed phase from the first and second
input wells of the unit to the channel junction, at which droplets of the dispersed phase are generated, and through the output
channel for collection in the output well of the unit.

US Pat. No. 9,132,394

SYSTEM FOR DETECTION OF SPACED DROPLETS

Bio-Rad Laboratories, Inc...

1. A detection system for droplet-based assays, comprising:
a channel network defining a flow path for droplets extending through a confluence region configured to increase an average
distance between droplets, and to an examination region disposed downstream of the confluence region; and

a detector operatively connected to the examination region,
wherein the channel network includes a droplet inlet channel configured to carry droplets to the confluence region, wherein
the droplet inlet channel forms a tapered region and a neck region extending from the tapered region to the confluence region,
wherein the tapered region is sized such that droplets enter the neck region in single file, wherein the flow path has a smaller
diameter in the neck region than in the examination region, and wherein at least one dilution inlet channel meets the droplet
inlet channel at the confluence region and is configured to provide a dilution fluid that increases the average distance between
droplets.

US Pat. No. 9,409,174

MICROFLUIDIC SYSTEM WITH FLUID PICKUPS

Bio-Rad Laboratories, Inc...

20. A system for fluid processing, comprising:
at least one well component providing an input well and an output well;
a channel component including a plurality of substantially horizontal microchannels that meet one another at a channel intersection,
the channel component having a bottom side that is affixed to and abutted with the at least one well component, and also having
a substantially vertical input tube projecting into the input well and disposed in fluid communication with the channel intersection;

at least one vacuum/pressure source operatively connected to the channel component; and
a source of carrier fluid disposed in fluid communication with the channel intersection;
wherein the system is configured to receive a sample-containing fluid in the input well such that the sample-containing fluid
is in contact with a bottom end of the input tube and is retained, with assistance from gravity, out of the plurality of microchannels
until a pressure differential is created with the at least one vacuum/pressure source that drives at least a portion of the
sample-containing fluid from the input well via the input tube to the channel intersection, and that drives at least a portion
of the carrier fluid to the channel intersection, such that droplets including the sample-containing fluid and disposed in
the carrier fluid are formed at the channel intersection and collected in the output well.

US Pat. No. 9,328,376

SYSTEMS AND METHODS FOR STABILIZING DROPLETS

Bio-Rad Laboratories, Inc...

1. A method for forming a droplet containing a sample to be detected, comprising:
a. generating a droplet at an intersection of a first channel and a second channel, said first channel in fluid communication
with a carrier fluid reservoir and said second channel in fluid communication with a sample reservoir, wherein said droplet
is generated by bringing a carrier fluid from said carrier fluid reservoir in contact with a sample or sample partition from
said sample reservoir at said intersection;

b. flowing said droplet along a droplet channel leading from said intersection to a droplet reservoir; and
c. heating said droplet such that a skin is formed around the droplet by protein denaturation before the droplet enters the
droplet reservoir.

US Pat. No. 9,200,318

REDUCED INHIBITION OF ONE-STEP RT-PCR

BIO-RAD LABORATORIES, INC...

1. A method of reverse transcribing a cDNA molecule from an RNA template, comprising:
mixing the RNA template with a composition comprising a reverse transcriptase, a DNA primer, and a phosphorothioate oligodeoxynucleotide,
wherein the phosphorothioate oligodeoxynucleotide is present in an amount from 0.01 nM to 25 nM; and

incubating the mixture at a temperature sufficient to synthesize a DNA molecule complementary to at least a portion of the
RNA template, thereby reverse transcribing the cDNA molecule.

US Pat. No. 9,945,866

PROTEIN STANDARD

Bio-Rad Laboratories, Inc...

1. A protein standard for gel electrophoresis, the standard comprising multiple protein sets, each set corresponding to a different gel band, wherein at least one set comprises a mixture of unlabeled and labeled proteins, said unlabeled protein comprising a tryptophan residue, and said labeled protein comprising a label that is spectrally distinct from haloalkylated tryptophan.

US Pat. No. 9,320,988

DEGASSING OF A LIQUID TO CONTROLLED LEVEL IN COMPOSITE TUBE

BIO-RAD LABORATORIES, INC...

1. A method of debubbling or degassing a liquid, the method comprising,providing:
a) a tube comprising a hollow lumen, a gas-permeable, liquid-impermeable wall surrounding said lumen, an upstream end, and
a downstream end;

b) a pump connected to the upstream end of the tube;
c) a device connected to the downstream end of the tube; andpassing the liquid through the lumen of the tube;wherein the pump drives the liquid into the tube, the device impedes the flow of the liquid out of the tube, and gasses or
bubbles escape through the gas-permeable, liquid-impermeable wall of the tube.

US Pat. No. 9,309,282

SOLID PHASE FOR MIXED-MODE CHROMATOGRAPHIC PURIFICATION OF PROTEINS

Bio-Rad Laboratories, Inc...

9. A method for purifying monomeric antibodies from a source solution comprising monomeric antibodies and antibody aggregates,
said method comprising:
(a) contacting said source solution at a pH of 4.0 to 6.0 with a mixed-mode chromatography medium comprising a ligand coupled
to a solid support, said ligand comprising benzamidoacetic acid, said solid support having pores of a median diameter of 0.5
micron or greater with substantially no pores of 0.1 micron or less in diameter, and said ligand coupled to said solid support
at a phenyl ring through a chain of one to three atoms, to bind said monomeric antibodies and antibody aggregates in said
source solution to said solid support through said ligand; and

(b) eluting said monomeric antibodies so bound from said solid support while said antibody aggregates remain bound to said
solid support, thereby purifying monomeric antibodies from a source solution comprising monomeric antibodies and antibody
aggregates.

US Pat. No. 9,442,891

MULTI-STAGE, REGRESSION-BASED PCR ANALYSIS SYSTEM

Bio-Rad Laboratories, Inc...

1. A method of determining a baseline region for an amplification curve resulting from a PCR amplification process of a biological
and/or chemical reaction sample, the method comprising:
detecting, with a detector of a polymerase chain reaction (PCR) system, fluorescence intensity values from the biological
and/or chemical reaction sample undergoing the PCR amplification process, the fluorescence intensity values corresponding
to a set of data points representing an amplification curve having a baseline portion and a growth portion, each data point
representing a fluorescence intensity value during the PCR amplification process;

receiving, at a computer system communicably coupled with the detector, the set of data points representing the amplification
curve having the baseline portion and the growth portion;

determining, by the computer system, a beginning of the baseline region, wherein determining the beginning of the baseline
region comprises:

determining whether an increase in a value of a one data point to a value of a next data point is greater than a predetermined
amount; and

if the predetermined amount is exceeded, selecting the beginning of the baseline region to be after the one data point;
determining, by the computer system, an end of the baseline portion; and
determining, by the computer system, a quantity of a target molecule in the biological and/or chemical reaction sample by
determining a Ct value using fluorescence intensity values in the determined baseline region and fluorescence intensity values in the growth
portion.

US Pat. No. 9,395,370

PURIFICATION OF MONOCLONAL ANTIBODIES

Bio-Rad Laboratories, Inc...

1. A method of identifying a pair of monoclonal antibodies for use in a sandwich immunoassay prior to the establishment of
individual hybridoma clones, the method comprising in the following order,
(a) contacting an array of non-clonal different hybridoma culture supernatants to an array of antibody-binding molecules,
wherein the antibody-binding molecules are linked to a solid support, under conditions to allow for binding of antibodies
in the supernatant to the antibody-binding molecule, wherein hybridomas in the hybridoma cultures were not previously isolated
to form clonal lines;

(b) separating unbound components of the supernatants from the support;
(c) altering the conditions of solution surrounding the solid support, thereby eluting polyclonal antibodies from the antibody-binding
molecules,

(d) separating the solid support from the solution comprising the eluted polyclonal antibodies of step (c) to create an array
of elutions comprising antibodies from the hybridomas; and

(e) determining the ability of a first antibody in the array of elutions to bind to an antigen in the presence of a second
antibody that binds the antigen, thereby identifying a pair of monoclonal antibodies for use in a sandwich immunoassay.

US Pat. No. 9,358,542

MAGNETIC TUBE RACK

Bio-Rad Laboratories, Inc...

1. A system for holding sample tubes, the system comprising:
a top plate defining a first plurality of holes, each of the first plurality of holes being a through hole through the top
plate and sized for accommodating an outer diameter of a respective sample tube;

a bottom plate defining a second plurality of holes, each of the second plurality of holes sized to accommodate a tip of a
respective sample tube, wherein the first and second pluralities of holes are positioned to cooperatively hold the sample
tubes in a pair of parallel rows;

a pair of end plates to which the top and bottom plates are attached at opposing ends, and that hold the top and bottom plates
in spaced-apart relation; and

a removable magnet holder configured to slide between the top plate and the bottom plate and between the two rows of sample
tubes, the magnet holder holding a plurality of magnets that, when the magnet holder is fully inserted between the rows of
sample tubes, align with the sample tubes in the parallel rows;

wherein a first of the two endplates defines an opening through which the magnet holder slides when the magnet holder is inserted
between the rows of sample tubes.

US Pat. No. 9,234,874

DIMENSIONAL STABILIZATION OF SLAB GEL CASSETTES TO PREVENT DISTORTION CAUSED BY SWELLING GELS

Bio-Rad Laboratories, Inc...

1. A device for insertion between a pair of parallel flat casting plates to form a row of sample wells in an electrophoresis
slab gel as said gel is cast between said plates, said device comprising:
a flat bar having a forward face, a rear face, and a longitudinal edge,
a row of dentiform projections along said longitudinal edge,
one or more first hook(s) projecting forward from said forward face, and
one or more second hook(s) projecting rearward from said rear face, said first and second hook(s) shaped to hold said casting
plates parallel to each other and at a fixed distance from each other during expansion of the gel with said dentiform projections
between said casting plates.

US Pat. No. 9,182,886

CHROMATOGRAPHY CONFIGURATION INTERFACE

Bio-Rad Laboratories Inc....

1. A graphical user interface generated by a processor and displayed on an electronic display screen for selecting from a
plurality of available process configurations, the graphical user interface comprising:
a plurality of adjacent category regions for displaying representing sections of a process
for each of the category regions, a plurality of dynamically selectable items representing different types of a respective
component appropriate for placement in the respective represented section of the process being configured, and a control that
causes selection of one of the selectable items and placement of the selected item into the respective one of the category
regions;

wherein the set of items currently selected in the plurality of category regions defines a selected process configuration.
US Pat. No. 9,453,208

NUCLEIC ACID MODIFYING ENZYMES

Bio-Rad Laboratories, Inc...

1. A nucleic acid encoding a polypeptide comprising two joined heterologous domains:
a sequence non-specific double-stranded nucleic acid binding domain that comprises an amino acid sequence that has at least
75% sequence identity to SEQ ID NO:2; or that comprises an amino acid sequence that has at least 75% sequence identity to
the Sac7d sequence set forth in amino acids 7-71 of SEQ ID NO:10; and

a DNA polymerase domain;
wherein the presence of the sequence non-specific double-stranded nucleic acid binding domain enhances the processivity of
the polymerase domain compared to an identical polymerase domain that does not have the sequence non-specific double-stranded
nucleic acid binding domain joined thereto.

US Pat. No. 9,388,945

SYSTEM FOR EMULSION ASPIRATION

Bio-Rad Laboratories, Inc...

1. A device for forming and holding an emulsion to be at least partially aspirated into a tip of a fluid-aspiration device,
the tip having a flat end surrounding an inlet, the device comprising:
a sample input channel;
one or more carrier input channels;
a droplet output channel;
a droplet generation region configured to form droplets of an emulsion, the droplet generation region being created where
the sample input channel, the one or more carrier input channels, and the droplet output channel meet one another; and

an outlet well fluidically connected to the droplet generation region via the droplet output channel, the outlet well including
a floor having one or more surface features that prevent uninterrupted circumferential contact of the flat end of the tip
with the floor.

US Pat. No. 9,383,354

ANTI-ANTIBODY REAGENT

BIO-RAD LABORATORIES, INC...

1. A panel of reagents comprising:
a binding member that binds an analyte;
the analyte coupled to a solid support; and
a mixture comprising (i) an unlabeled antibody that binds the binding member and (ii) a labeled antibody that binds the binding
member at an unlabeled:labeled antibody ratio of 2:1 to about 20:1.

US Pat. No. 9,347,059

METHODS AND COMPOSITIONS FOR NUCLEIC ACID ANALYSIS

BIO-RAD LABORATORIES, INC...

1. A method comprising:
a. generating a plurality of first partitions comprising adaptors, wherein each of the first partitions has on average a first
volume and wherein the adaptors comprise unique barcodes, wherein the first partitions are first droplets in a fluid that
is immiscible to the first droplets;

b. generating a plurality of second partitions comprising sample polynucleotides, wherein each of the second partitions has
on average a second volume, wherein the second volume is greater than the first volume, wherein the second partitions are
second droplets in a immiscible fluid that is immiscible to the second droplets;

c. applying a treatment to fuse the at least one second partition with at least one first droplet to form a fused partition;
and

d. tagging one of the sample polynucleotides, or fragment thereof, with at least one of the adaptors in the fused partition
to form tagged polynucleotides or fragments thereof.

US Pat. No. 9,347,094

DIGITAL ASSAY FOR TELOMERE LENGTH

Bio-Rad Laboratories, Inc...

1. A method of characterizing telomere length, the method comprising:
synthesizing a template with a first primer and a second primer in a bulk phase reaction mixture containing genomic DNA, the
template representing a repetitive telomeric sequence present in the genomic DNA and containing a binding site introduced
by one of the primers, wherein the one primer includes a 3? portion that binds a repetitive telomeric sequence in the genomic
DNA and also includes a 5? portion that is not required for binding to the repetitive telomeric sequence, and wherein the
5? portion of the one primer introduces the binding site;

forming partitions after synthesizing the template in the bulk phase reaction mixture, with only a subset of the partitions
containing at least one copy of the template;

amplifying at least a region of the template in partitions to form an amplicon in the presence of a probe that specifically
binds to a region of the amplicon that corresponds to the binding site;

collecting amplification data from the probe in partitions; and
determining a measure of telomere length for the genomic DNA based on the amplification data.

US Pat. No. 9,310,600

DOUBLE FOLD OPTICS

Bio-Rad Laboratories, Inc...

1. An imaging assembly comprising:
a platform having a target region;
a camera and lens module;
a reflex mirror, positioned along an optical path incident from the camera and lens module, wherein the reflex mirror is oriented
relative to the camera and lens module with a yaw angle of about 15° to about 35°; and

a focal plane mirror, positioned along an optical path incident from the reflex mirror, wherein the focal plane mirror is
oriented relative to the reflex mirror with a pitch angle of about 15° to about 25°, the optical path between the camera and
lens module and the target region being non-orthogonal, having an imaging distance length of about 50 cm to about 70 cm, and
having a focal plane in the target region.

US Pat. No. 9,442,050

REDUCING PH EXCURSIONS IN ION EXCHANGE CHROMATOGRAPHY USING DISPLACING COUNTER IONS

BIO-RAD LABORATORIES, INC...

1. A method of suppressing formation of pH excursions on a charged solid support having associated hydrogen or hydroxide ions,
the method comprising
providing a solid support in association with an equilibration buffer, a wash buffer, or a loading buffer and subsequently;
contacting the solid support with a solution comprising displacing counter ions with a charge opposite the net charge of the
solid support, wherein the displacing counter ion is a positively-charged molecule thereby replacing hydrogen ions associated
with a negatively charged solid support, or wherein the displacing counter ion is a negatively-charge molecule thereby replacing
hydroxide ions associated with a positively charged solid support; and subsequently

eluting a target molecule from the solid support, wherein the eluting comprises increasing conductivity of an elution solution
in contact with the solid support.

US Pat. No. 9,399,215

SAMPLE HOLDER WITH A WELL HAVING A WICKING PROMOTER

Bio-Rad Laboratories, Inc...

1. A device for holding a sample, comprising:
a holder including an upper member attached to a lower member to form a well having a side wall region and a floor, the side
wall region extending from a top border to the floor and widening toward the floor at a position intermediate the top border
and the floor to create a distinct edge that conceptually divides the well into an upper portion and a lower portion, the
well including a side wall protrusion defined by the upper member and adapted to promote wicking of an aqueous sample into
the lower portion from the upper portion, the holder defining a channel communicating with the lower portion of the well and
with a channel junction that provides a droplet generator;

wherein the side wall protrusion is contiguous with the edge.

US Pat. No. 9,156,010

DROPLET-BASED ASSAY SYSTEM

Bio-Rad Laboratories, Inc...

1. A system for sample analysis, comprising:
a plurality of droplet generators that form a plurality of separate emulsions including droplets of partitioned samples, each
droplet generator including a channel intersection at which one of the emulsions is formed from at least one carrier fluid
stream and a sample stream, the samples being prepared as reaction mixtures for amplification of at least one nucleic acid
target;

containers defining cavities to hold the emulsions;
a transport structure to transfer the emulsions from the droplet generators to the containers;
a heating and cooling device to heat the emulsions in the cavities, with the emulsions held in an array, to induce nucleic
acid amplification in droplets of the emulsions;

a detection assembly to detect signals from intact droplets of the emulsions;
a controller in communication with the detection assembly and programmed to estimate a presence, if any, of the nucleic acid
target in the samples based on signals detected from the intact droplets;

at least one pressure source that drives flow of the at least one carrier fluid stream and the sample stream to the channel
intersection of each droplet generator; and

a transport structure to transfer droplets of the emulsions from the containers to the detection assembly.
US Pat. No. 9,470,655

POLYACRYLAMIDE GELS FOR RAPID CASTING, BLOTTING, AND IMAGING, WITH STORAGE STABILITY

Bio-Rad Laboratories, Inc...

1. A kit for forming a polyacrylamide gel, the kit comprising:
a first container comprising a solution comprising acrylamide and bis-acrylamide; and
a second container comprising triethanolamine, an ampholyte selected from the group consisting of glycine and tricine, and
a conjugate ampholyte consisting of an amino acid with a pKa within the range of 8.3 to 9.6.

US Pat. No. 9,371,167

ANTI-COLLAPSE FLEXIBLE FLUID CONTAINER

BIO-RAD LABORATORIES, INC...

1. A valve assembly for holding chemistry assay fluid within a flexible container, the valve assembly comprising:
a rigid container having a first opening;
a flexible container having a second opening, the flexible container being mounted within the rigid container such that first
opening and second opening are in alignment;

a valve anchored in the second opening;
an extended beam support structure, extending from the valve within the interior of the flexible container, the extended beam
support structure adapted to prevent an interior surface of the flexible container from blocking fluidic communication between
the interior of the flexible container and the valve and further comprising at least one crosspiece having flow openings positioned
along the length of the extended beam support structure.

US Pat. No. 9,200,305

GENERATION OF ENGINEERED MOLECULAR WEIGHT STANDARDS

Bio-Rad Laboratories, Inc...

1. A molecular weight marker composition that comprises an engineered polypeptide that comprises an amino acid sequence having
at least 80% identity to an amino acid sequence set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:8; wherein
the protein characterized by the amino acid sequence retains the same charge and molecular weight as a reference protein set
forth in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7.

US Pat. No. 9,383,311

NON-SCANNING SPR SYSTEM

Bio-Rad Laboratories Inc....

14. A method of measuring reflectivity of light from a surface exhibiting an evanescent wave phenomenon at total internal
reflection, the method comprising:
a) reflecting light from a plurality of sensing areas of interest which exhibit the evanescent wave phenomenon, arranged one-dimensionally
on a surface, over a range of angles of incidence for each area, one or more of the sensing areas of interest being capable
of binding to material in a fluid sample that comes in contact with them;

b) projecting an image of the surface to a plurality of secondary optical elements, with sufficiently sharp focus so that
light reflected from different sensing areas of interest is projected to substantially different secondary elements;

c) projecting the light from the secondary elements to a detector sufficiently close to a focal plane of the secondary elements
so that light reflected from each sensing area of interest at different angles of incidence within its range of angles is
spread out to sufficiently different locations on reaching the detector to distinguish the detector response for the different
angles of incidence; and

d) determining, from a response of the detector, how much light is reflected from each sensing area of interest, as a function
of angle of incidence over the range of angles for that area, for at least one time interval.

US Pat. No. 9,322,002

SSO7-POLYMERASE CONJUGATES WITH DECREASED NON-SPECIFIC ACTIVITY

Bio-Rad Laboratories, Inc...

1. An Sso7 polymerase conjugate protein comprising an Sso7 domain linked to a polymerase; wherein:
the Sso7 domain is at least 75% identical to SEQ ID NO:2;
an amino acid of the Sso7 domain corresponding to R43 of SEQ ID NO:2 is not arginine (R); and
the conjugate protein has a decreased non-specific amplification activity compared to an otherwise identical control conjugate
protein in which the amino acid of the Sso7 domain corresponding to R43 of SEQ ID NO:2 is arginine (R).

US Pat. No. 9,500,664

DROPLET GENERATION FOR DROPLET-BASED ASSAYS

Bio-Rad Laboratories, Inc...

14. A method of manufacturing a droplet generation system, comprising:
(i) forming a substrate having a bottom surface and a top surface;
(ii) forming a sample well;
(iii) forming a background fluid well;
(iv) forming a droplet well; and
(v) forming a droplet generation region defined by the intersection of a first channel fluidically connected with the sample
well, a second channel fluidically connected with the background fluid well, and a third channel fluidically connected with
the droplet outlet region;

wherein the substrate, an upper region of the sample well, an upper region of the background fluid well, and an upper region
of the droplet well are injection molded as a single piece;

wherein the upper region of each well protrudes from the top surface of the substrate; and
wherein the first channel, the second channel, and the third channel are formed in the bottom surface of the substrate.
US Pat. No. 9,354,144

CUSTOMIZED QUALITY CONTROLS FOR ANALYTICAL ASSAYS

Bio-Rad Laboratories, Inc...

1. A method for preparing a set of controls for an assay of one or more selected analytes in a sample of human biological
fluid by a selected assay technique, said assay to determine a concentration of each of said analytes in said sample relative
to a known medical decision point concentration for that analyte, said method comprising:
dissolving a plurality of quantities of one or more solid water-soluble beads in separate aliquots of an aqueous liquid matrix,
the aliquots having equal volumes of the aqueous liquid matrix,

wherein each said water-soluble bead is prepared by lyophilization of an aqueous solution ranging in volume from about 5 ?L
to about 1,000 ?L and comprises at least one said analyte, a bulking agent, a salt, and a buffer, and

said aqueous liquid matrix comprising a salt and a buffer at a pH of from about 4.0 to about 9.0,
to form a plurality of liquid solutions having a range of concentrations for each said analyte, the range bracketing said
medical decision point concentration for that analyte,

such that the concentration of each of said analytes is less than or equal to said medical decision point concentration in
at least one liquid solution, and the concentration of each of said analytes is greater than or equal to said medical decision
point concentration in at least one liquid solution.

US Pat. No. 9,139,873

METHODS OF USING IMPROVED POLYMERASES

Bio-Rad Laboratories, Inc...

1. A method of sequencing a target nucleic acid in an aqueous solution using an improved DNA polymerase, the method comprising:
(a) contacting the target nucleic acid with a thermally stable DNA polymerase, wherein the DNA polymerase is joined to a sequence
non-specific double-stranded nucleic acid binding domain that comprises at least 75% amino acid sequence identity to the Sso7d
amino acid sequence set forth in SEQ ID NO:2, and enhances the processivity of the DNA polymerase compared to an identical
DNA polymerase not having the sequence non-specific double-stranded nucleic acid binding domain fused to it, and

wherein the solution is of a composition that permits the sequence non-specific double-stranded nucleic acid binding domain
to bind to the target nucleic acid and the polymerase domain to extend a primer that is hybridized to the target nucleic acid
sequence;

(b) incubating the solution under conditions in which the primer is extended by the DNA polymerase; and
(c) identifying the nucleotides that are incorporated into the synthesized nucleic acid strand upon the template-dependent
extension of the primer by the DNA polymerase, thereby sequencing the nucleic acid.

US Pat. No. 9,486,741

HYDROLYSIS-RESISTANT POLYACRYLAMIDE GELS

Bio-Rad Laboratories, Inc...

1. A polyacrylamide gel comprising
crosslinked polyacrylamide,
triethanolamine, and
a catalytic amount of a polymerization catalyst selected from the group consisting of ammonium persulfate, N,N?-tetramethylenediamine
(TEMED), riboflavin, ?-dimethylamino-propionitrile, and combinations thereof;

said gel having
(i) from about 0.03 mM to about 0.3 mM of tris(hydroxymethyl)aminomethane, and having from about 0.03 mM to about 0.3 mM of
bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane; or

(ii) from about 0.03 mM to about 0.3 mM of tris(hydroxymethyl)aminomethane, and being devoid of bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane;
or

(iii) from about 0.03 mM to about 0.3 mM of bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, and being devoid of tris(hydroxymethyl)aminomethane;
wherein the polyacrylamide gel has a pH of from about 6.4 to about 7.0, and is configured to separate and resolve proteins
under electrophoretic conditions.

US Pat. No. 9,417,190

CALIBRATIONS AND CONTROLS FOR DROPLET-BASED ASSAYS

Bio-Rad Laboratories, Inc...

1. A method of performing a droplet-based assay, comprising:
forming droplets from a same mixture, wherein only a subset of the droplets contain a target;
irradiating a region of a channel;
detecting a forward scattering signal and a fluorescence signal from a plurality of the droplets as each droplet passes through
the region of the channel, wherein the fluorescence signal is detected from a label present in each droplet and varies according
to whether the target is present in the droplet;

identifying accepted droplets of the plurality for which the forward scattering signal meets a condition and rejected droplets
of the plurality for which the forward scattering signal does not meet the condition, wherein the condition corresponds to
a permitted size of a droplet; and

determining a concentration of the target based on the fluorescence signal from the accepted droplets, and without any contribution
of the fluorescence signal from the rejected droplets.

US Pat. No. 9,347,095

DIGITAL ASSAYS FOR MUTATION DETECTION

Bio-Rad Laboratories, Inc...

1. A method for detecting a first and a second allele of a target locus in a plurality of target polynucleotide molecules,
comprising:
(a) partitioning a sample comprising said target polynucleotide molecules into a plurality of fluid reaction volumes;
(b) performing an amplification reaction in said reaction volumes, wherein each reaction volume comprises (i) a forward primer
that is complementary to a first sequence of a first strand of the target polynucleotide molecules, wherein the first sequence
is 5? of the target locus, (ii) a reverse primer that is complementary to a second sequence of a second strand of the target
polynucleotide molecules, wherein the second sequence is 3? of the target locus, and (iii) a detection probe that is capable
of hybridizing to a third sequence of the target polynucleotide molecules that encompasses the target locus, wherein the detection
probe comprises a first signal reporter; and

(c) determining a number and/or fraction of positive reaction volumes containing a target molecule having the first allele
and a number and/or fraction of reaction volumes containing a target molecule having the second allele based on a signal of
the first signal reporter detected from the reaction volumes.

US Pat. No. 9,393,560

DROPLET TRANSPORT SYSTEM FOR DETECTION

Bio-Rad Laboratories, Inc...

1. A method of transporting droplets for detection, comprising:
providing an emulsion disposed in a container and including droplets;
creating contact between a tip and the emulsion by moving at least one of the tip and the container relative to each other,
the tip being connected to an examination region and including an outer tube and an inner tube, the outer tube forming a first
open end and surrounding an enclosed portion of the inner tube, the inner tube extending out of the first open end to create
a projecting portion forming a second open end below the first open end;

loading droplets of the emulsion into the inner tube via the second open end;
moving loaded droplets from the inner tube to the examination region; and
dispensing a first fluid onto the projecting portion of the inner tube from the first open end formed by the outer tube, and
a second fluid from the second open end formed by the inner tube.

US Pat. No. 9,327,213

PREPARATIVE CHROMATOGRAPHY COLUMN

Bio-Rad Laboratories, Inc...

1. A chromatography column comprising:
a rigid column shell,
a bottom plate secured to said rigid column shell and comprising a layer of porous material covering a rigid and liquid-impermeable
bottom plate base wherein said rigid and liquid-impermeable bottom plate base has a bottom plate port therein for passage
of liquid,

a piston comprising a filter covering a rigid and liquid-impermeable piston base and said rigid and liquid-impermeable piston
base having a piston port for passage of liquid, said piston fitting within a tube within said rigid column shell, and

said tube being of flexible, water-impermeable material containing and in contact with a separation medium, said tube being
open at a first end such that the tube is in full contact with the rigid column shell, closed at a second end with said bottom
plate, and encircling said piston.

US Pat. No. 9,274,103

CONJUGATES OF 1,4,7-TRIAZACYCLONONANES, DINUCLEAR METAL COMPLEXES OF SUCH CONJUGATES, AND METHODS OF USE FOR BOTH 1,4,7-TRIAZACYCLONONANES AND CONJUGATES

Bio-Rad Laboratories, Inc...

1. A method of staining phosphorylated compounds immobilized on a solid support or in a gel with a fluorescent dye, said method
comprising contacting said solid support or gel with a compound having the formula
in which:
one of R1 through R6 is -L-R7 in which L is a member selected from the group consisting of —CH2—C(?O)—NH—(C1-C4 alkyl)-, —CH2—C(?O)—NH—(C1-C4 alkyl)-NH—, and —(C1-C4 alkyl)-NH—, and R7 is a fluorescent dye;

the remainder of R1 through R6 are independently selected from the group consisting of H and C1-C6 alkyl; and

M is a divalent metal.
US Pat. No. 9,212,203

SURFACE NEUTRALIZATION OF APATITE

BIO-RAD LABORATORIES, INC...

1. A method for purifying a target molecule in a sample, the method comprising,
(a) equilibrating an apatite solid surface with a buffer composition suitable to adsorb the target molecule on the apatite
solid surface and then contacting the sample comprising the target molecule to the apatite solid surface thereby adsorbing
the target molecule to the solid surface;

(b) after (a), contacting the solid surface comprising the adsorbed target molecule with a solution comprising:
(i) a basic amino compound and an alkali metal ion; or
(ii) a sulphonated amine compound and an alkali metal ion,
of sufficient volume and concentration to neutralize the apatite solid surface wherein the solution has a sufficiently low
ionic strength such that the target molecule remains adsorbed to the solid support, wherein the buffer composition in (a)
is of different composition from the solution of (b); and

(c) after (b), eluting the target molecule from the solid support by contacting the solid support with a solution of different
composition from the solution in (b), thereby purifying the target molecule in the sample.

US Pat. No. 9,243,288

CARTRIDGE WITH LYSIS CHAMBER AND DROPLET GENERATOR

Bio-Rad Laboratories, Inc...

1. A method of nucleic acid amplification, comprising:
purifying a fluid sample in a portion of a disposable, single-use cartridge;
lysing the sample in a portion of the cartridge;
combining the purified, lysed sample with a reagent mixture in a portion of the cartridge;
driving flow of at least one carrier fluid stream and a sample stream to a channel intersection at which droplets containing
the sample are generated to form an emulsion, wherein the sample stream includes the purified, lysed sample combined with
the reagent mixture; and

cycling the emulsion thermally in a thermocycling instrument to which the emulsion is transferred from the cartridge,
wherein the step of combining includes a step of advancing a first plunger that is operatively connected to a first reservoir
containing one of the sample and the reagent mixture, while retracting a second plunger that is operatively connected to a
second reservoir containing the other of the sample and the reagent mixture, to move fluid out of the first reservoir and
into the second reservoir.

US Pat. No. 9,217,175

MULTIPLEXED DIGITAL ASSAY WITH SPECIFIC AND GENERIC REPORTERS

Bio-Rad Laboratories, Inc...

1. A method of performing a multiplexed digital assay, the method comprising:
providing partitions each including a portion of a same mixture, the mixture containing a first target and a second target
and also containing a generic reporter that is sensitive to amplification of either target and a specific reporter that is
specifically sensitive to amplification of the second target, wherein only a first subset of the partitions each contain at
least one copy of the first target and only a distinct second subset of the partitions each contain at least one copy of the
second target;

amplifying the first target and the second target in the partitions;
collecting amplification data from the generic reporter and the specific reporter present in a plurality of the partitions;
and

calculating a level of each target based on the amplification data.
US Pat. No. 9,493,824

UNIVERSAL REFERENCE DYE FOR QUANTITATIVE AMPLIFICATION

Bio-Rad Laboratories, Inc...

1. A reaction mixture for signal normalization in a real-time polymerase chain reaction (PCR) amplification of a target nucleic
acid wherein the mixture is compatible, without further addition of reagents aside from a sample to be tested and optionally
primers, for use in both (a) a high concentration passive dye normalization real-time PCR amplification system and (b) a low
concentration passive dye normalization real-time PCR amplification system, wherein the mixture comprises a plurality of passive
reference dyes that produces fluorescent signals independent of the amplification reactions.

US Pat. No. 9,921,154

MULTIPLEXED DIGITAL ASSAYS

Bio-Rad Laboratories, Inc...

1. A method of performing a multiplexed digital assay, the method comprising:
forming partitions that collectively contain R targets;
amplifying the R targets in the partitions;
collecting data representing amplification of each of the R targets in the partitions, all of the data being collected in
fewer than R optical channels; and

determining a respective level of each of the R targets from the data, wherein each level is specific for a single target
of the R targets, and wherein the level determined for at least one of the R targets is based in part on a partition count
for a partition population positive for two of the R targets;

wherein the step of determining is based on at least R+2 identified partition populations.

US Pat. No. 9,945,808

INSTRUMENT FOR INDEPENDENT ELECTROTRANSFER IN MULTIPLE CASSETTES

BIO-RAD LABORATORIES, INC...

1. An electrotransfer instrument comprising:a housing with an internal cavity containing a plurality of electroblotting cassettes in a vertical stack;
a power supply;
individual electrical contacts mounted to said internal cavity of said housing for connecting said power supply via the electrical contacts to electrodes in said cassette that is inserted therein; and
electrical relays configured to engage said electrical contacts for each one of said plurality of cassettes upon insertion of said one cassette, and configured to block said power supply to said electrical contacts upon removal of said one cassette.

US Pat. No. 9,945,809

DRY PROTEIN TRANSFER

Bio-Rad Laboratories, Inc...

1. A kit for transferring biological macromolecules from an electrophoresis slab gel to a blotting membrane by electroblotting, the kit comprising two conductive polymer electrodes, wherein each electrode comprises:a matrix comprising a conductive polymer blend, the conductive polymer blend comprising a conjugated organic polymer, and
a terminal electrically coupled to the matrix, wherein the terminal is configured to receive electrical power from a power supply,
wherein:
each electrode further comprises a rigid cassette encasing the matrix, the cassette comprising a floor, walls connected to the floor, and a lip extending laterally from the walls;
the floor and walls of the cassette define a central cavity; and
the matrix is disposed in the central cavity, and
either of:
(a) the terminal of each electrode is disposed on an external surface of the cassette and is electrically coupled to the matrix through a conductive member, the conductive member passing through a wall of the cassette or the floor of the cassette; or
(b) a plurality of alignment pegs protrude from the lip of the cassette of at least one electrode,
a plurality of alignment holes are cut into the lip of the cassette of at least one electrode,
the alignment pegs and alignment holes are positioned to engage each other when the electrodes are brought together, and
the alignment holes are complementary in shape to the alignment pegs, so that engagement of the alignment pegs with the alignment holes secures the electrodes together while leaving a gap between the matrices.

US Pat. No. 9,850,517

COMPACT AUTOMATED CELL COUNTER

Bio-Rad Laboratories, Inc...

1. A sample slide for counting cells in a cell suspension, said sample slide comprising:
a flat plate with an internal chamber bounded by optically clear upper and lower windows, wherein the internal chamber has
at least one overflow area positioned at a corner of the internal chamber, wherein the at least one overflow area is open
at the top of said sample slide, wherein the internal chamber has a loading port and a vent port, and wherein the loading
port and the vent port are at two opposing longitudinal ends of the internal chamber; and

a first and a second orientation notch in said flat plate, wherein the first and second orientation notches are in diagonally
opposing corners of said flat plate, and wherein the first and second orientation notches guide said flat plate into a cell
counting instrument in an orientation whereby said upper window faces a selected direction within said instrument.

US Pat. No. 9,606,111

STAIN-FREE PROTEIN QUANTIFICATION AND NORMALIZATION

Bio-Rad Laboratories, Inc...

1. A method of quantifying a total amount of protein in a complex target protein sample by detected haloalkylated tryptophan
fluorescence, said method comprising:
a) providing a complex target protein sample and a plurality of complex reference protein samples;
b) exposing said complex target protein sample of step a) and said plurality of complex reference protein samples of step
a) to UV light while said complex target protein sample and said complex reference protein samples are in contact with a haloalkane,
thereby covalently reacting tryptophan residues of proteins in said complex target protein sample and said complex reference
protein samples with said haloalkane, and

c) detecting fluorescence emitted by said proteins in said complex target protein sample of step b) and said plurality of
complex reference protein samples of step b), wherein said fluorescence is haloalkylated tryptophan fluorescence;

d) providing a total amount of protein in said plurality of complex reference protein samples of step c), and quantifying
an amount of haloalkylated tryptophan fluorescence detected from said plurality of complex reference protein samples of step
c);

e) quantifying an amount of haloalkylated tryptophan fluorescence detected from said complex target protein sample of step
c); and

f) quantifying said total amount of protein in said complex target protein sample based upon said total amounts of protein
in said plurality of complex reference protein samples of step d) and said amounts of haloalkylated tryptophan fluorescence
detected from said plurality of complex reference protein samples of step d) and said complex target protein sample of step
e),
wherein said complex target protein sample comprises 50 or more proteins and said plurality of complex reference protein samples
comprise 50 or more proteins.

US Pat. No. 9,222,128

MULTIPLEXED DIGITAL ASSAYS WITH COMBINATORIAL USE OF SIGNALS

Bio-Rad Laboratories, Inc...

1. A method of performing a multiplexed digital amplification assay, the method comprising:
amplifying more than R targets in partitions;
creating R signals representative of light detected in R different wavelength regimes from the partitions, where R?2; and
calculating an average level of each target in the partitions based on the R signals, wherein the level calculated accounts
for a coincidence of all possible combinations of the more than R targets in the same individual partitions;

wherein the more than R targets include three targets, and wherein the average levels of the three targets are calculated
based on light detected from only two fluorophores associated with probes that bind to amplicons of the three targets during
amplification.

US Pat. No. 10,048,256

SAMPLE ANALYSIS SYSTEMS AND METHODS

Bio-Rad Laboratories, Inc...

1. A method of determining the presence or absence of an analyte in a sample, the method comprising:applying a substance to a surface of a substrate having a first binding agent immobilized thereon, wherein the first binding agent is capable of binding to the substance;
removing unbound material from at least a portion of the substrate having the substance applied thereon;
applying a second binding agent to the surface of the substrate, wherein the second binding agent is optically labeled or unlabeled;
removing unbound material from at least a portion of the substrate having the second binding agent applied thereon; and either
responsive to detecting the second binding agent bound to the substance, identifying the analyte present in the sample; or
responsive to not detecting the second binding agent bound to the substance, determining that the analyte is absent in the sample,
wherein the applying the substance or second binding agent to the surface of the substrate steps are concurrent with the respective removing unbound material from at least a portion of the substrate steps.
US Pat. No. 9,127,042

ENHANCED PURIFICATION OF ANTIBODIES AND ANTIBODY FRAGMENTS BY APATITE CHROMATOGRAPHY

Bio-Rad Laboratories, Inc...

1. A method for purifying at least one non-aggregated antibody or at least one immunoreactive antibody fragment from an impure
preparation containing said antibody or antibody fragment comprising the steps of (a) contacting the impure preparation with
an apatite chromatography support and (b) conducting elution in the presence of an ionic species selected from the group consisting
of proprionate, pyruvate, gluconate, glucuronate, proline, lysine, and histidine.

US Pat. No. 9,126,160

SYSTEM FOR FORMING AN ARRAY OF EMULSIONS

Bio-Rad Laboratories, Inc...

1. A system for forming an array of emulsions in parallel, comprising:
a plate providing an array of emulsion production units each configured to produce a separate emulsion and each including
a set of wells interconnected by a set of channels forming a channel junction, each channel being bounded circumferentially,
each set of wells including

at least one first input well to receive a continuous phase,
a second input well to receive a dispersed phase, and
an output well;
wherein the set of channels includes at least two input channels extending separately from the input wells to the channel
junction, at which droplets of the dispersed phase are generated in the continuous phase, and an output channel extending
from the channel junction to the output well, in which an emulsion is collected.

US Pat. No. 9,507,809

SYSTEM AND METHOD FOR PROVIDING AUTOMATICALLY UPDATED PRODUCT INSERTS

Bio-Rad Laboratories, Inc...

1. A method of distributing updated parameters of operating results for one or more control products used in biological reactions,
the method comprising:
receiving, from a user by a server system, a request to customize delivery of one or more update notifications for one or
more control products, wherein the one or more update notifications are to be sent when one or more parameters associated
with the one or more control products have been updated since sending a previous update notification to the user;

accessing, by the server system, a database to determine whether the one or more parameters have been updated since the previous
update notification, each test operable to test one or more analytes in a biological reaction to generate one or more analyte
results and operable to test the one or more control products in the biological reaction to generate one or more control results,
the one or more control results operable to confirm a quality of the one or more analyte results, wherein the request identifies
the one or more analytes and the one or more control products;

when the one or more parameters have been updated since the previous update notification, receiving, by the server system
from the database, data that has been updated since the previous update notification, wherein the data comprises one or more
of the following: a date or time stamp of the updated parameters, a flag identifying a change from the previous update notification,
and the one or more parameters; and

sending by the server system, the update notification to the user, wherein the update notification includes at least part
of the data that has been updated since receiving a previous request.

US Pat. No. 9,683,792

FLOATING THERMAL CONTACT ENABLED PCR

Bio-Rad Laboratories, Inc...

1. A thermal management device enabling PCR reactions in a microfluidic channel, the device comprising:
a framework;
one or more thermal zone sub-assemblies coupled to the framework, each thermal zone sub-assembly having a thermal control
element, and wherein each thermal zone sub-assembly is coupled to the framework by one or more thermal actuation mechanisms;
and

one or more thermal spreaders configured to contact the thermal control elements of the one or more thermal zone sub-assemblies,
wherein the thermal actuation mechanisms consist essentially of insulative bearings, fasteners, and a spring, wherein the
spring is configured to apply a force to drive the thermal control element toward one of the one or more thermal spreaders,
wherein each fastener is a shoulder screw, and wherein each thermal actuation mechanism includes at least one polymer-based
bearing, which travels up and down on the shaft of at least one of the shoulder screws that are attached to the framework
and are pushed by the spring.

US Pat. No. 9,145,550

COMPOSITIONS WITH POLYMERASE ACTIVITY

Bio-Rad Laboratories, Inc...

1. A polymerase that comprises a hybrid polymerase domain that has at least 94% identity to the amino acid sequence set forth
in SEQ ID NO:12; or the polymerase region of SEQ ID NO:6, SEQ ID NO:8, or SEQ ID NO:10, wherein the hybrid polymerase domain
has an increased ratio of polymerase activity to exonuclease activity relative to Pyrococcus sp. GD-B polymerase when joined to a sequence non-specific double-stranded nucleic acid binding domain.
US Pat. No. 9,134,301

SORTING OF ADHERENT CELLS BY SELECTIVE TRANSFORMATION OF LABELS

Bio-Rad Laboratories, Inc...

1. A method for sorting a population of biological cells to detect a selected subpopulation thereof having a characteristic
not shared by other cells of said population, said method comprising:
(a) while cells of said population are in adherent form,
(i) labeling cells of said subpopulation and cells not of said subpopulation with a transformable label that is capable of
undergoing a transformation upon exposure to energy, the transformation resulting in a detectable difference in the label
as compared with a label that has not been exposed to said energy, wherein the detectable difference is retained upon converting
labeled cells from adherent form to nonadherent form,

(ii) identifying individual cells of said population belonging to said subpopulation or not belonging to said subpopulation,
and

(iii) specifically exposing the individual cells so identified to label-transforming energy to transform labels on cells so
exposed;

(b) subsequently converting cells in adherent form of said population to nonadherent form; and
(c) while cells of said population are in nonadherent form, sorting said population to distinguish between cells of said subpopulation
and cells other than said subpopulation by differentiating between cells with labels so transformed and cells with labels
that have not been so transformed.

US Pat. No. 9,176,154

CALIBRATION PROCESS AND SYSTEM

Bio-Rad Laboratories, Inc...

1. A method of performing a calibration in a medical testing machine, the method comprising:
automatically detecting that a package of calibration material has been inserted into the medical testing machine;
performing, by the medical testing machine, a calibration sequence using the calibration material to ascertain a calibration
parameter for the medical testing machine; and

storing the calibration parameter in electronic storage for use in later tests performed by the medical testing machine;
wherein the calibration material is provided in a lyophilized state, the method further comprising controlling the medical
testing machine to automatically reconstitute the lyophilized material before performing the calibration sequence.

US Pat. No. 9,162,229

MULTI-DIRECTIONAL SORTING WITH REDUCED CONTAMINATION IN A FLOW CYTOMETER

Bio-Rad Laboratories, Inc...

1. A collection sled for a flow cytometer comprising:
a first sample collector located in a first position to collect samples from a first deflected droplet stream in said flow
cytometer, said first sample collector having apertures that accept standard size sample collection tubes;

a second sample collector located in a second position to collect samples from a second deflected droplet stream in said flow
cytometer, said second sample collector having apertures that accept standard size sample collection tubes, said second sample
collector spaced apart from said first sample collector to form a trough between said first sample collector and said second
sample collector along a side portion of said first sample collector and a side portion of said second sample collector, said
trough being aligned with said waste droplet stream so that said waste droplet stream passes through said trough into a cavity
formed by said first sample collector and said second sample collector;

a waste stream collector formed from said cavity between said first sample collector and said second sample collector that
expands in a downward direction from said trough to prevent backsplash of waste fluid from waste droplets in said waste droplet
stream and has a waste disposal port for disposing of waste fluid from said waste droplet stream.

US Pat. No. 9,983,205

MICROFLUIDIC DEVICES FOR AUTOMATED ASSAYS

Bio-Rad Laboratories, Inc...

1. A cartridge comprising:a block frame comprising:
(a) a well, wherein the well comprises an outlet, at least one inlet, and a bottom surface;
(b) a plurality of containers embedded in the block frame, wherein each container is connected to the well via a microchannel leading to the at least one inlet; and
(c) a trough separate from and surrounding the well, said trough configured to hold a liquid; and
an openable cover, which cover when closed is configured to enclose an assay surface and form a gap between (i) the assay surface and the bottom surface of the well or (ii) the assay surface and the cover.

US Pat. No. 9,764,322

SYSTEM FOR GENERATING DROPLETS WITH PRESSURE MONITORING

Bio-Rad Laboratories, Inc...

1. A method of generating droplets, the method comprising:
selecting a device including a plurality of emulsion-formation units each including a sample well, a continuous-phase well,
a droplet well, and a channel network that fluidically interconnects the wells and creates a droplet-generation region;

placing a discrete volume of sample-containing fluid into the sample well of each emulsion-formation unit, and a discrete
volume of continuous-phase fluid into the continuous-phase well of each emulsion-formation unit;

applying pressure to the device with a fluidics assembly after the step of placing, such that in each emulsion-formation unit
(a) sample-containing fluid flows from the sample well to the droplet-generation region, (b) continuous-phase fluid flows
from the continuous-phase well to the droplet-generation region, and (c) sample-containing droplets and continuous-phase fluid
flow from the droplet-generation region to the droplet well, wherein the plurality of emulsion-formation units generate droplets
in parallel with one another;

detecting a pressure signal from the fluidics assembly; and
stopping application of the pressure when the pressure signal indicates that a sample well is empty.

US Pat. No. 9,489,251

SUPERVISING AND RECOVERING SOFTWARE COMPONENTS ASSOCIATED WITH MEDICAL DIAGNOSTICS INSTRUMENTS

BIO-RAD LABORATORIES, INC...

1. A system for applying a recovery mechanism to a plurality of computer software components that control medical diagnostics,
the system comprising:
a plurality of medical diagnostics instruments, each associated with a computer software component;
a plurality of communication modules, each associated with a corresponding one of the plurality of the computer software components,
wherein each one of the plurality of communication modules is arranged to report on malfunctioning computer software components;
and

a recovery module, configured to:
(i) periodically obtain reports from the communication modules every certain period of time, the reports enabling a determination
of whether computer software components are malfunctioning;

(ii) reload the malfunctioning computer software components, wherein reloading the malfunctioning computer software component
of a first medical diagnostics instrument comprises creating, at a communication layer, a client for providing a service to
another computer software component of a second medical diagnostic instrument; and

(iii) notify all communication modules of the reloading of the malfunctioning computer software components;
wherein each communication module is further configured to re-establish a connection between the corresponding one of the
plurality of the computer software components and the reloaded computer software components.

US Pat. No. 9,248,417

SYSTEM FOR DROPLET-BASED ASSAYS USING AN ARRAY OF EMULSIONS

Bio-Rad Laboratories, Inc...

1. A system for performing a droplet-based assay, comprising:
a plate including a plurality of emulsion production units;
an instrument configured to be operatively connected to the plate to drive production of emulsions collected in wells of the
plate;

a droplet transporter configured to pick up droplets from the emulsions held in an array of wells and to drive flow of the
droplets through a detection region, wherein each well holds an emulsion including a plurality of droplets;

a detector configured to collect data related to one or more analytes from individual droplets of the emulsions as such individual
droplets travel through the detection region; and

a controller programmed to determine, based on the data collected, an aspect of the one or more analytes in one or more samples
included in droplets of the emulsions.

US Pat. No. 9,149,809

THERMAL CYCLER WITH VAPOR CHAMBER FOR RAPID TEMPERATURE CHANGES

BIO-RAD LABORATORIES, INC...

1. An apparatus for thermal cycling in an array of sample receptacles, said apparatus comprising:
a hollow body having a single internal cavity with a working fluid therein that is partially vaporized, wherein said hollow
body has a top surface with depressions therein that are spaced and shaped to receive a plurality of sample receptacles and
thereby to place said depressions in direct and continuous contact with said undersides of said sample receptacles, said hollow
body arranged to cause conductive heat transfer between walls of said cavity and walls of all of said sample receptacles,
and wherein said internal cavity of said hollow body has a substantially flat floor and said depressions have undersides internal
to said cavity that contact said flat floor; and

one or more element that controllably heats said working fluid, cools said working fluid, or both, within said cavity to cause
vaporization and condensation of said working fluid, wherein said element is below or at a side of the hollow body.

US Pat. No. 9,909,959

CUSTOMIZED QUALITY CONTROLS FOR ANALYTICAL ASSAYS

Bio-Rad Laboratories, Inc...

1. A method for determining linearity of an assay response, said method comprising:
dissolving one or more solid water-soluble beads in three or more aliquots of an aqueous liquid matrix to prepare three or
more liquid solutions each having a different analyte level,

wherein the water-soluble beads comprise at least one analyte, a bulking agent, a salt, and a buffer, and
said aqueous liquid matrix comprising a salt and a buffer at a pH of from about 4.0 to about 9.0,
performing the assay with the three or more liquid solutions to generate an assay response for the analyte in each of the
three or more liquid solutions; and

determining whether assay responses are linear within a range of analyte concentrations spanned by three or more liquid solutions,
thereby determining linearity of an assay response.

US Pat. No. 9,861,979

INJECTION OF MULTIPLE VOLUMES INTO OR OUT OF DROPLETS

Bio-Rad Laboratories, Inc...

1. A system for injecting multiple volumes into droplets, comprising
at least one microfluidic channel intersected by two or more injection channels, wherein each injection channel forms an injection
inlet at an injection interface where each injection channel intersects the microfluidic channel, wherein the microfluidic
channel and injection interfaces are configured such that droplets within the microfluidic channel can be flowed from a first
location to a second location in the microfluidic channel past the injection interfaces and

one single pair of electrodes positioned to disrupt interfaces between droplets and a fluid and/or emulsion at the injection
inlets of the two or more injection channels,

wherein the at least one microfluidic channel comprises one or more droplets flowing therein, and wherein each of the two
or more injection channels comprises at least one fluid and/or emulsion therein.

US Pat. No. 9,649,635

SYSTEM FOR GENERATING DROPLETS WITH PUSH-BACK TO REMOVE OIL

Bio-Rad Laboratories, Inc...

23. A method of forming and concentrating an emulsion, the method comprising:
driving sample-containing fluid in a first channel and continuous-phase fluid in a second channel to a droplet-generation
region in which the first and second channels meet one another, such that sample-containing droplets are formed;

driving sample-containing droplets and continuous-phase fluid from the droplet-generation region to a well via a third channel,
such that an emulsion including sample-containing droplets disposed in continuous-phase fluid is collected in the well; and

decreasing a volume fraction of continuous-phase fluid in the emulsion by selectively driving continuous-phase fluid, relative
to the droplets, from the well via the third channel.

US Pat. No. 9,458,484

REVERSE TRANSCRIPTASE MIXTURES WITH IMPROVED STORAGE STABILITY

Bio-Rad Laboratories, Inc...

1. A frozen sterile aqueous reaction mixture comprising sufficient ingredients, except template RNA, for reverse transcription
of RNA, the mixture comprising:
a reverse transcriptase having an optimal pH above 8;
deoxytriphosphate nucleotides; and
a buffer, wherein the buffer is selected from the group consisting of Tris, HEPES, ACES, PIPES, MOPSO, BES, MOPS, TES, TAPSO,
POPSO, BICINE, TAPS, and AMPSO,

wherein said frozen sterile aqueous reaction mixture has a pH of between 6-8 and lacks template RNA.
US Pat. No. 9,856,525

DIGITAL ASSAYS WITH ASSOCIATED TARGETS

Bio-Rad Laboratories, Inc...

1. A method of analysis, the method comprising:
creating a mixture containing
(a) a sample comprising nucleic acid isolated from a first type of biological particle and a second type of biological particle,
wherein the first type of biological particle provides a first target and a second target having a covalent linkage to one
another in the isolated nucleic acid, and wherein the second type of biological particle contains the first target but not
the second target,

(b) reagents for amplification of the first target and the second target, and
(c) one or more reporters;
forming droplets each containing a portion of the mixture;
amplifying the first target and the second target in the droplets;
detecting at least one signal from the one or more reporters present in individual droplets;
classifying individual droplets as positive or negative for each target based on the at least one signal;
determining a first number of droplets that are positive for both targets, or a first number of droplets that are negative
for at least one of the targets;

determining a second number of droplets that are positive for the first target and negative for the second target, or a second
number of droplets that are negative for the first target and positive for the second target; and

determining a first level of the first type of biological particle using the first number and a second level of the second
type of biological particle using the second number.

US Pat. No. 9,791,408

MODIFIED ELECTRODE BUFFERS FOR STAIN-FREE PROTEIN DETECTION IN ELECTROPHORESIS

Bio-Rad Laboratories, Inc...

1. A kit comprising a first electrode buffer, wherein the first electrode buffer comprises a first halo-substituted organic
compound and a first buffering agent, and the first halo-substituted organic compound bears a charge in a system in which
the pH is equal to the pKa of the first buffering agent, wherein
the first electrode buffer is a cathode buffer, and the first halo-substituted organic compound bears a negative charge in
a system in which the pH is equal to the pKa of the first buffering agent, wherein the first halo-substituted organic compound
is tribromoacetate or trichloropropanoate or a hydrophobic compund encapsulated in a negatively charged micelle; or

the first electrode buffer is an anode buffer, and the first halo-substituted organic compound bears a positive charge in
a system in which the pH is equal to the pKa of the first buffering agent.

US Pat. No. 9,636,682

SYSTEM FOR GENERATING DROPLETS—INSTRUMENTS AND CASSETTE

Bio-Rad Laboratories, Inc...

1. A system for generating droplets, comprising:
a device including a sample well configured to receive sample-containing fluid, a continuous-phase well configured to receive
continuous-phase fluid, and a droplet well, the device also including a channel network having a first channel, a second channel,
and a third channel that meet one another in a droplet-generation region;

a holder for the device; and
an instrument configured to operatively receive an assembly including the device and the holder and to drive sample-containing
fluid from the sample well to the droplet-generation region via the first channel, continuous-phase fluid from the continuous-phase
well to the droplet-generation region via the second channel, and sample-containing droplets from the droplet-generation region
to the droplet well via the third channel.

US Pat. No. 9,304,518

MODULAR AUTOMATED CHROMATOGRAPHY SYSTEM

Bio-Rad Laboratories, Inc...

1. A system for joining a plurality of fluid manipulation components into a flow scheme for directing fluids to and from a
chromatographic separation device and for operating said joined fluid manipulation components according to a selected protocol,
said system comprising:
a plurality of modules, each said module comprising (a) one of said fluid manipulation components and (b) a microcontroller
that (i) encodes machine readable data characterizing the type of said fluid manipulation component, and (ii) transmits operational
signals to said fluid manipulation component in response to commands received from outside said module;

a mounting frame comprising a plurality of mounting sites, each said mounting site constructed to receive a single module
interchangeably with other said modules, and each said mounting site comprising a signal connector that couples to and communicates
with the microcontroller of a module mounted at said mounting site; and

a software platform embedded in said mounting frame or in a computer external to said mounting frame or distributed between
said mounting frame and a computer external to said mounting frame, the software communicating with the microcontroller of
each module mounted on said mounting frame through said signal connectors, said software being programmable for (1) receiving
said machine-readable data, (2) recognizing from the received machine-readable data the types of the fluid manipulation components,
(3) automatically mapping the plurality of fluid manipulation components to the flow scheme based on the received data, (4)
guiding a user of the system in the joining of said fluid manipulation components according to said flow scheme, and for (5)
causing said fluid manipulation components, once joined, to direct fluids to and from said chromatographic separation device
in accordance with said protocol.

US Pat. No. 10,152,519

OPTIMIZED SPECTRAL MATCHING AND DISPLAY

Bio-Rad Laboratories, Inc...

1. A method for identifying one or more substances in a sample, the method comprising performing, by a computer system:shining a light beam on the sample;
measuring a sample spectrum of the sample by detecting light transmitted or reflected by the sample using a detector, the sample spectrum having an intensity value for each of a plurality of wavelengths;
for each of a plurality of reference substances:
retrieving, from a database, a reference spectrum for the respective reference substance, the reference spectrum having intensity values for the plurality of wavelengths;
initially selecting one or more correction values for one or more correction parameters to be applied to at least one of the sample spectrum and the reference spectrum, the one or more correction parameters corresponding to at least one of a clipping correction, a horizontal shift correction, a vertical offset correction, or a baseline correction;
for each of a plurality of iterations:
applying the one or more correction values for the one or more correction parameters to at least one of the sample spectrum and the reference spectrum;
computing a similarity score between the sample spectrum and the reference spectrum resulting from application of the one or more correction values, the similarity score determined using differences between the intensity values of the reference spectrum and corresponding intensity values of the sample spectrum;
determining whether the similarity score satisfies one or more convergence criteria;
upon determining that the similarity score satisfies the one or more convergence criteria, identifying the similarity score as an optimized similarity score corresponding to one or more optimized values of the one or more correction parameters; and
upon determining that the similarity score does not satisfy the one or more convergence criteria, updating the one or more correction values for use in performing another iteration until the similarity score satisfies the one or more convergence criteria; and
comparing the optimized similarity score to a threshold to determine whether the sample contains the reference substance; and
outputting data about one or more of the plurality of reference substances that have optimized similarity scores that are above the threshold.

US Pat. No. 9,285,359

PROTEIN DETECTION USING MODIFIED CYCLODEXTRINS

BIO-RAD LABORATORIES, INC...

1. A method of detecting a protein, the method comprising:
contacting the protein with a cyclodextrin covalently linked to at least one label;
contacting the label with a reagent;
causing the reagent to undergo a chemical interaction with the at least one label to form a product;
detecting the product, thereby detecting the protein.
US Pat. No. 9,150,611

AQUEOUS TRANSFER BUFFER

Bio-Rad Laboratories, Inc...

1. A method for electrophoretic transfer comprising the step of transferring polypeptides of about 200 kDa to about 6.5 kDa
in molecular weight from a first substrate to a second substrate in an electrical field after the first substrate and the
second substrate are placed in an aqueous solution comprising Tris at a concentration of about 300 mM and glycine at a concentration
of about 300 mM.

US Pat. No. 9,068,916

MICROASSEMBLED IMAGING FLOW CYTOMETER

Bio-Rad Laboratories, Inc...

1. A microassembled imaging cytometer, comprising:
a sensing location that undergoes relative motion with a cell;
a light source that produces a beam;
a focusing element, the focusing element focusing light from the beam of light to a plurality of focused illumination spots
at a focal plane at the sensing location, such that the cell is illuminated by one or more of the focused illumination spots
as the cell traverses the sensing location, wherein each of the focused illumination spots is smaller than the cell;

an array light sensor that comprises an array of pixels and produces signals indicating the intensity and distribution of
light falling on the pixels;

a collection lens that collects and refocuses light emanating from the cell to a plurality of separate, discernible spots
on the array light sensor; and

a processing unit that constructs a digital image of the cell based at least in part on the signals.

US Pat. No. 10,033,794

NETWORK TRANSFER OF LARGE FILES IN UNSTABLE NETWORK ENVIRONMENTS

Bio-Rad Laboratories, Inc...

1. A method comprising performing, by a server computer:receiving, from a client computer, a first request to establish a first HTTP connection;
receiving, over the first HTTP connection, metadata for a file as part of the first request, the metadata including at least a filename, a client ID, and a file size;
determining that at least a portion of the file is not stored at the server computer based on the filename;
sending, to the client computer over the first HTTP connection, an indication of the at least a portion of the file to be sent by the client computer to the server computer;
sending, over the first HTTP connection, a suggested chunk size to be utilized by the client computer when sending the at least a portion of the file to the server computer;
receiving, over the first HTTP connection, one or more chunks of the file, each chunk having a size less than or equal to the suggested chunk size, from the client computer, and each chunk specifying a start index and an end index for the chunk;
storing information indicating parts of the file that have been received;
losing the first HTTP connection between the server computer and the client computer;
receiving, from the client computer, a second request to establish a second HTTP connection;
receiving, over the second HTTP connection, the metadata for the file as part of the second request;
determining that the file has not been completely received;
sending a missing start index and a missing end index corresponding to a missing part of the file to the client computer;
receiving the missing part of the file from the client computer; and
sending a confirmation that the file has been stored completely to the client computer.

US Pat. No. 9,864,832

SNP DETECTION BY MELT CURVE CLUSTERING

Bio-Rad Laboratories, Inc...

1. A method of identifying a sequence variation between nucleotide sequences, the method comprising:
subjecting each sample of a set of samples to a Polymerase Chain Reaction (PCR) growth process in a PCR system to produce
copies of a double stranded molecule of two nucleotide sequences in each sample;

gradually increasing a temperature of the PCR system such that the double-stranded PCR products melt into single strands;
detecting, with the PCR system, a plurality of sets of data points, each set of data points corresponding to a different sample,
each data point of a set of data points including a signal value and a temperature value for the sample where the temperature
value increases for each successive data point, wherein each set of data points defines a melt curve;

determining a melt region having a melt region start and a melt region end;
assigning each melt curve to a respective cluster, wherein the melt curves assigned to a same cluster have one or more similar
shape properties in the melt region relative to melt curves in other clusters;

at least one processor selecting a cluster of melt curves;
the at least one processor determining a melting temperature of each melt curve of the selected cluster;
the at least one processor grouping the melt curves of the selected cluster into a plurality of sub-clusters based on the
respective melting temperatures;

identifying at least a portion of the nucleotide sequences corresponding to at least one sub-cluster as having a sequence
variation relative to the nucleotide sequences of another sub-cluster; and

subjecting a sample corresponding to the identified portion of the nucleotide sequences to sequencing to determine the sequence
variation.

US Pat. No. 9,829,434

OPTICAL DETECTION SYSTEM FOR LIQUID SAMPLES

Bio-Rad Laboratories Inc....

1. A tip for use in an optical detection system to analyze an analyte in a fluid sample drawn into the tip, using light reflected
from a detection surface inside the tip that the analyte binds to, comprising:
at least one active surface having a surface chemistry with at least one functional group that allows it to form a covalent
bond with at least one amine group of a ligand; and

at least one reference surface having a surface chemistry that do not form a covalent bond with said at least one amine group
of said ligand;

wherein said functional groups of said active surface and said reference surface have similar non-specific binding properties;
wherein said at least one functional group of said at least one active surface comprises a carboxylic group; and
wherein said at least one functional group of said at least one reference surface comprises at least one functional group
selected from the group consisting of sulfate, sulfonate, and phosphonate groups.

US Pat. No. 9,745,571

REPETITIVE REVERSE TRANSCRIPTION PARTITION ASSAY

Bio-Rad Laboratories, Inc...

1. A method of quantifying one or more target RNA(s) in a biological sample, the method comprising,
incubating a plurality of mixture partitions of the sample, wherein the mixture partitions are droplets, and wherein the mixture
comprises the sample and a heat tolerant reverse transcriptase, under conditions such that at least two and up to 20 rounds
of reverse transcription occurs in the partitions, wherein the rounds of reverse transcription are each terminated by a denaturation
event, and wherein the denaturation event comprises heating the partition mixtures to a temperature above 55° C., thereby
generating one or more cDNA(s) complementary to at least 10 contiguous nucleotides of the target RNA in partitions having
the target RNA;

quantifying the number of partitions containing the cDNA(s); and
correlating the number of partitions containing the cDNA(s) to the quantity of the one or more target RNA(s) in the sample,
thereby quantifying the one or more target RNA(s) in the sample.

US Pat. No. 9,669,404

ANTI-COLLAPSE FLEXIBLE FLUID CONTAINER

Bio-Rad Laboratories, Inc...

1. A valve assembly for holding chemistry assay fluid within a flexible container, the valve assembly comprising:
a rigid container having a first opening;
a flexible container having a second opening, the flexible container being mounted within the rigid container such that first
opening and second opening are in alignment;

a valve anchored in the second opening;
a corkscrew-shaped cage support structure, extending from the valve within the interior of the flexible container, the support
structure adapted to prevent an interior surface of the flexible container from blocking fluidic communication between the
interior of the flexible container and the valve.

US Pat. No. 9,492,797

SYSTEM FOR DETECTION OF SPACED DROPLETS

Bio-Rad Laboratories, Inc...

1. A method of detection for droplet-based assays, the method comprising:
placing an open end of a channel network into an emulsion;
driving droplets of the emulsion along a flow path from the open end, through a confluence region where a dilution fluid is
introduced into the flow path from at least one dilution inlet channel to increase an average distance between droplets, and
through an examination region disposed downstream of the confluence region; and

detecting light from the examination region as droplets pass through;
wherein the channel network includes a droplet inlet channel that meets the at least one dilution inlet channel at the confluence
region, and wherein the droplet inlet channel forms a tapered region that is sized such that droplets leave the tapered region
in single file.

US Pat. No. 9,089,844

SYSTEM FOR FORMING EMULSIONS

Bio-Rad Laboratories, Inc...

1. A system for emulsion formation, comprising:
a microfluidic device having a plurality of emulsion formation units each including a sample well, a droplet well, a sample
inlet channel extending from the sample well to a channel intersection, and a droplet outlet channel extending from the channel
intersection to the droplet well; and

an instrument that operatively receives the microfluidic device and including a fluidics assembly having a pressure sensor,
the instrument being configured (a) to apply pressure to the emulsion formation units in parallel with the fluidics assembly
to drive parallel generation of droplets at the channel intersections of the emulsion formation units and parallel collection
of emulsions of the droplets in the droplet wells of the emulsion formation units, (b) to monitor the pressure with the pressure
sensor, and (c) to stop application of the pressure to all of the emulsion formation units when the pressure sensor detects
a change in pressure indicative of air entering any one of the sample inlet channels from a corresponding sample well.

US Pat. No. 9,091,654

SERIAL-LINE-SCAN-ENCODED MULTI-COLOR FLUORESCENCE MICROSCOPY AND IMAGING FLOW CYTOMETRY

Bio-Rad Laboratories, Inc...

1. A system for performing cytometry, the system comprising:
a scanning region that is illuminated by light including at least first and second wavelength bands;
a cell transport mechanism transporting a cell through the scanning region such that the cell is illuminated;
a set comprising at least one linear light sensor; and
an optical system that selectively directs light emitted from the cell to two portions of the linear light sensor set such
that emitted light in a third wavelength band is primarily directed to a first portion of the linear light sensor set, and
emitted light in a fourth wavelength band is primarily directed to a second portion of the linear light sensor set;

wherein the system repeatedly takes readings of light falling on the linear light sensor set while the cell is transported
through the scanning region.

US Pat. No. 9,605,060

MULTIPLEX IMMUNOASSAYS FOR HEMOGLOBIN, HEMOGLOBIN VARIANTS, AND GLYCATED FORMS

BIO-RAD LABORATORIES, INC...

2. A monoclonal antibody that selectively binds to hemoglobin variant HbE and glycated HbE, wherein the antibody binds to
a HbE minimal epitope 22EVGGK26 (SEQ ID NO:6) or 21DEVGGK26 (SEQ ID NO:7); and is raised against an HbE immunogen consisting of H2N-CYG-VTALWGKVNVDEVGGK-CONH2 (SEQ ID NO:29).
US Pat. No. 9,556,475

AMPLIFICATION REPORTER WITH BASE-PAIRING OLIGOMERS

Bio-Rad Laboratories, Inc...

1. A method of analysis, the method comprising:
forming at least one volume containing a reporter including a first oligomer and a second oligomer capable of base-pairing
with one another below a melting temperature of the reporter to affect a photoluminescence detectable from a photoluminophore
of the first oligomer, wherein the first oligomer and the second oligomer are not covalently linked to one another;

amplifying a target in the at least one volume, at least in part by extending one or more primers at a temperature above the
melting temperature of the reporter, wherein the step of amplifying causes cleavage of the first oligomer, and wherein the
cleavage affects the photoluminescence detectable from the photoluminophore;

detecting photoluminescence from the photoluminophore in the at least one volume while the at least one volume is at a temperature
below the melting temperature of the reporter; and

determining a level of the target based on the photoluminescence detected.

US Pat. No. 9,205,379

INSTRUMENT FOR INDEPENDENT ELECTROTRANSFER IN MULTIPLE CASSETTES

Bio-Rad Laboratories, Inc...

9. A method for transferring electrophoretically separated species from a slab gel to a sheet-form matrix, said method comprising:
(a) placing said slab gel and a sheet-form matrix in an electroblotting cassette that comprises anode and cathode plates and
external electrical contacts for each of said plates;

(b) inserting said cassette in the electrotransfer instrument of claim 1 such that said external electrical contacts are in contact with said individual electrical contacts; and
(c) imposing electrical charges on said electrodes to cause said species to migrate electrophoretically from said slab gel
to said sheet-form matrix within said inserted cassette.

US Pat. No. 9,110,050

MEASURING MULTI-ANALYTE SAMPLES USING AN IN-LINE FLOW CELL

Bio-Rad Laboratories, Inc...

1. A method for measuring analytes in a sample, the method comprising:
flowing a sample containing analytes through a detector;
measuring a measurand of the analytes passing through the detector;
determining a time interval over which the measurand reaches a predetermined quantity of mass, wherein measuring the measurand
comprises performing a real-time integration of the measurand over the time interval;

forming a sample plug based on the time interval over which the measurand reaches the predetermined quantity of mass; and
diverting a portion of the flowing sample containing the sample plug to a sample volume.

US Pat. No. 9,821,312

INTEGRATED MICROFLUIDIC SYSTEM, METHOD AND KIT FOR PERFORMING ASSAYS

Bio-Rad Laboratories, Inc...

1. A method for forming assay droplets comprising the steps of:
providing a microfluidic device comprising a first plurality of reagent droplets, the first plurality of reagent droplets
comprising a first reagent set, and a second plurality of reagent droplets, the second plurality of droplets comprising a
second reagent set;

forming, from a sample and the first plurality of reagent droplets, a first plurality of assay droplets in the microfluidic
device, wherein the forming comprises injecting in the microfluidic device at least a portion of a sample into some or all
of the first plurality of reagent droplets; and

forming, from the first plurality of assay droplets and the second plurality of reagent droplets, a second plurality of assay
droplets, wherein the forming comprises injecting in the microfluidic device different portions of an assay droplet from the
first plurality of assay droplets into multiple droplets from the second plurality of reagent droplets, thereby creating a
second plurality of assay droplets in the microfluidic device.

US Pat. No. 9,766,213

INNOVATION TO ASSAY MIXING

Bio-Rad Laboratories, Inc...

1. A method of calibrating a fluid delivery system, the method comprising:
providing a fluid delivery system connected to N input streams, wherein N is an integer greater than or equal to two, and
wherein the fluid delivery system mixes together fluids from the N input streams to form an output stream;

obtaining N test solutions, wherein each test solution comprises a dye having a characteristic wavelength of maximum absorbance
(?max);

for each of the N test solutions,
designating a wavelength ?test, wherein ?test is within the range from 90% of ?max to 110% of ?max, and

providing an input absorbance of the test solution, the input absorbance being the absorbance of the solution at ?test;

injecting the N test solutions into the N input streams;
mixing the N test solutions in the fluid delivery system;
measuring the absorbance of the output stream at N wavelengths, the wavelengths being the ?tests designated for the test solutions, thereby obtaining an output absorbance for each test solution; and

comparing the input absorbance with the output absorbance for each test solution, thereby calibrating the fluid delivery system.

US Pat. No. 9,726,871

DETECTION SYSTEM WITH ONE-PIECE OPTICAL ELEMENT TO CONCENTRATE AND HOMOGENIZE LIGHT

Bio-Rad Laboratories, Inc...

1. A method of sample examination, the method comprising:
disposing at least a portion of a sample in an examination region, wherein the at least a portion of a sample is disposed
in droplets, and wherein the droplets are disposed in a carrier phase;

illuminating the examination region such that the sample emits light;
concentrating emitted light from the examination region with a one-piece optical element;
homogenizing the intensity of the concentrated light with the optical element by internal reflection; and
detecting light of homogenized intensity received from the optical element.
US Pat. No. 9,599,543

CUSTOMIZED QUALITY CONTROLS FOR ANALYTICAL ASSAYS

BIO-RAD LABORATORIES, INC...

1. A kit for preparation of a customized control for a diagnostic assay, said kit comprising:
an aqueous liquid matrix comprising a salt and a buffer at a pH of from about 4.0 to about 9.0, wherein the aqueous liquid
matrix comprises human or animal source materials from which endogenous species that interfere with the detection of said
one or more selected analyte have been removed; and

a plurality of solid water-soluble beads, wherein the solid water-soluble beads are prepared by lyophilization of an aqueous
solution ranging in volume from about 5 ?L to about 1,000 ?L, and the solid water-soluble beads comprise at least one analyte,
a bulking agent, a salt, and a buffer,

wherein said beads are configured for 1) dissolving different quantities of the one or more solid water-soluble bead in separate
aliquots of said aqueous liquid matrix, wherein the aliquots have equal volume; or 2) dissolving the same quantities of the
one or more solid water-soluble bead in separate aliquots of said aqueous liquid matrix, wherein the aliquots have different
volume.

US Pat. No. 9,354,155

CELL COUNTING SYSTEMS AND METHODS

Bio-Rad Laboratories, Inc...

21. A method of counting cells, the method comprising:
capturing, using a sensing system, a first digital image of a first viewable portion of a sample containing cells;
analyzing, using a computerized analyzer, the first digital image to obtain a count of the cells present in the first digital
image;

comparing the count with a threshold count;
when the count is below the threshold count, reconfiguring the sensing system and sample such that the sensing system images
a second viewable portion of the sample;

capturing a second digital image of the second viewable portion; and analyzing the second digital image and reporting a test
result based on the count of the cells present in the first and second digital images; and

when the count is above the threshold count, reporting a test result based on a count of the cells present in the first digital
image without capturing a second digital image;

wherein the first and second viewable portions are arranged transversely with respect to a primary loading direction of the
sample into the sample holder;

wherein the count is a first count and the threshold count is a first threshold count, the method further comprising, when
the first count is below the first threshold count;

analyzing the first and second digital images to compute a second count of the cells present in the first and second digital
images;

comparing the second count with a second threshold count;
when the second count is below the second threshold count, reconfiguring the sensing system and sample such that the sensing
system images a third viewable portion of the sample; capturing a third digital image of the third viewable portion; and analyzing
the third digital image and reporting a test result based on a count of the cells present in the first, second, and third
digital images; and

when the second count is above the second threshold count, reporting a test result base on a count of the cells present in
the first and second digital images without capturing a third digital image.

US Pat. No. 9,193,760

DISSOCIATION OF PRODUCT-COMPLEXED CONTAMINANTS IN CHROMATOGRAPHY

Bio-Rad Laboratories, Inc...

1. A method of purifying a target molecule from a biological sample, the method comprising in the following order,
(a) contacting the sample comprising the target molecule and complexed contaminants to a metal cation-derivatized apatite
solid support or a polycation-derivatized apatite solid support, wherein the metal cation is calcium, thereby non-covalently
binding the target molecule to the solid support so as to provide a bound target molecule;

(b) washing the bound target molecule with an agent(s) that displaces the complexed contaminants from the target molecule
under conditions in which the target molecule remains substantially bound to the solid support, wherein the agent(s) comprise
urea, 0.5-2.0 M sodium chloride, or both; and

(c) eluting the target molecule from the solid support, wherein the eluted target molecule is substantially free of complexed
contaminants.

US Pat. No. 9,194,861

METHOD OF MIXING FLUIDS BY COALESCENCE OF MULTIPLE EMULSIONS

Bio-Rad Laboratories, Inc...

1. A method of sample analysis, comprising:
obtaining a plurality of compound droplets each including at least one sample-containing droplet, at least one reagent-containing
droplet, and a barrier fluid that separates the at least one sample-containing droplet from the at least one reagent-containing
droplet;

heating compound droplets to induce fusion of at least one of the sample-containing droplet(s) with at least one of the reagent-containing
droplet(s) within individual compound droplets to form fused droplets that combine sample and reagent to create assay mixtures
for performing an assay;

collecting data from a plurality of the fused droplets; and
determining a result of the assay based on the collected data.
US Pat. No. 9,127,312

ANALYSIS OF NUCLEIC ACIDS

Bio-Rad Laboratories, Inc...

1. A method of detecting variations in copy number of a target nucleic acid, the method comprising:
a. contacting a sample comprising a plurality of polynucleotides with at least one agent, wherein each of one or more of polynucleotides
comprises multiple copies of a target nucleic acid;

b. subjecting the sample to conditions that enable the agent to cleave a specifically-selected target site between the multiple
copies of the target nucleic acid, thereby separating the multiple copies from one another;

c. separating the sample contacted with the agent into a plurality of droplets;
d. subjecting the target nucleic acid to an amplification reaction in droplets;
e. detecting amplification data for the target nucleic acid from individual droplets;
f. enumerating a number of droplets comprising the target nucleic acid based on the amplification data; and
g. determining a copy number of the target nucleic acid based on the step of enumerating.
US Pat. No. 9,927,439

MULTIPLEX HEPATITIS B ASSAY

Bio-Rad Laboratories, Inc...

1. A kit comprising a Hepatitis B detection assay comprising detection of multiple Hepatitis B virus (HBV) antibodies in a subject sample comprising:(a) a first receptacle comprising a mixture of
1) beads covalently conjugated to human Hepatitis B surface antigen (HBsAg) and detectable label (i), wherein the conjugated beads bind antibodies that are specific for HBsAg, if present,
2) beads covalently conjugated to human Hepatitis B core antigen (HBcAg) and detectable label (ii), wherein the conjugated beads bind antibodies that are specific for HBcAg, if present, and
3) beads covalently conjugated to an antibody specific for human IgM (anti-IgM) and detectable label (iii), wherein the conjugated beads bind human IgM, if present;
(b) a second receptacle comprising
4) biotinylated HBsAg, and
5) biotinylated HBcAg; and
(c) a third receptacle comprising streptavidin covalently conjugated to detectable label (iv).

US Pat. No. 9,845,496

MULTIPLEXED DIGITAL ASSAY WITH SPECIFIC AND GENERIC REPORTERS

Bio-Rad Laboratories, Inc...

1. A composition for performing a multiplexed digital assay, comprising:
an emulsion including droplets, each droplet including a portion of a same aqueous phase and containing (i) a generic reporter
that is sensitive to amplification of a first target and a second target and (ii) a specific reporter that is sensitive to
amplification of the second target but not the first target;

wherein each droplet includes a complete set of reagents for amplification of each target, if present in the droplet,
wherein each droplet of a subset of the droplets contains the first target but not the second target, and
wherein each droplet of a subset of the droplets contains the second target but not the first target.
US Pat. No. 9,708,598

METHODS OF USING IMPROVED POLYMERASES

Bio-Rad Laboratories, Inc...

1. A method of increasing the yield from a polymerase reaction on a target nucleic acid present in a solution that comprises
a fluorescent dye that detects double-stranded DNA, the method comprising:
(a) contacting the target nucleic acid with a DNA polymerase, wherein the DNA polymerase comprises a polymerase domain fused
to a sequence-non-specific double-stranded nucleic-acid-binding domain that comprises at least 75% amino acid sequence identity
to the Sso7d amino acid sequence set forth in SEQ ID NO:2 and enhances the processivity of the DNA polymerase compared to
an identical DNA polymerase not having the sequence non-specific double-stranded nucleic-acid binding domain fused to it;

wherein the solution comprises the fluorescent dye and is of a composition that permits the sequence non-specific double-stranded
nucleic acid binding domain to bind to the target nucleic acid and the polymerase domain to extend a primer that is hybridized
to the target nucleic acid sequence;

(b) incubating the solution under conditions in which the primer is extended by the polymerase.

US Pat. No. 9,696,257

AUTOMATED AND ACCURATE DROP DELAY FOR FLOW CYTOMETRY

Bio-Rad Laboratories, Inc...

1. A flow cytometry system for selecting a desired drop delay for a flow cytometer, the flow cytometer comprising:
means for forming a stream such that the stream forms droplets at a droplet separation point;
means for interrogating the stream at an interrogation point upstream of the droplet separation point, wherein the desired
drop delay is representative of the elapsed time between when particles pass through the interrogation point and when a droplet
containing the particles separates from the stream at the droplet separation point;

means for charging the droplets;
means for electrostatically deflecting the droplets;
waste tubing configured to form a waste stream that is generated from droplets that are not deflected by the means for electrostatically
deflecting the droplets;

a waste stream optical source configured to illuminate the waste stream in the waste tubing;
a detector configured to sense scattered light or fluorescent emissions from calibration particles in the waste stream; and
a controller that is configured to:
operate the flow cytometer for a plurality of drop delay periods while calibration particles are present in the stream, each
drop delay period having a different selected drop delay from a range of potential selected drop delays,

cause the means for charging to charge the droplets during each drop delay period according to the selected drop delay for
that drop delay period,

cause the means for electrostatically deflecting the droplets to deflect the charged droplets during each of the drop delay
periods,

determine information indicative of a rate at which the calibration particles enter the waste stream during each drop delay
period based on the sensed scattered light or fluorescent emissions from the calibration particles in the waste stream, and

determine the desired drop delay based upon the information indicative of the rate at which the calibration particles enter
the waste stream during each drop delay period.

US Pat. No. 9,535,000

ON-DEMAND PARTICLE DISPENSING SYSTEM

Bio-Rad Laboratories, Inc...

1. A method of isolating a particle from a fluid sample, the method comprising:
streaming a fluid sample containing a particle through a sample channel; and
diverting the particle from the sample channel into an outlet channel, based on a detected characteristic of the particle,
by directing a diverting fluid into the sample channe upstream and downstream of the particle.

US Pat. No. 9,273,347

DETECTION OF RNA-INTERACTING REGIONS IN DNA

Bio-Rad Laboratories, Inc...

1. A method of detecting RNA-interacting regions in genomic DNA, the method comprising:
introducing an RNA-degrading agent and a DNA-degrading agent into a nucleus, whereby at least one DNA region in the genomic
DNA is degraded by the DNA-degrading agent due to the presence of the RNA-degrading agent;

detecting the at least one DNA region in the genomic DNA that is degraded by the DNA-degrading agent; and
comparing the amount of the at least one DNA region that is detected to a control sample in which the genomic DNA is contacted
by the DNA-degrading agent but not the RNA-degrading agent, wherein an absence or reduction in the quantity of copies of a
DNA region, relative to the control sample, indicates that the DNA region is degraded by the DNA-degrading agent; thereby
detecting the RNA-interacting regions.

US Pat. No. 9,058,648

IMAGE ACQUISITION FOR CHEMILUMINESCENT SAMPLES

Bio-Rad Laboratories, Inc...

1. A method for measuring the chemiluminescence of a sample, comprising:
acquiring a binned initial image of the sample, the initial image having a first binning value representing the number of
pixel elements in its image pixels;

calculating an exposure time for a final image of the sample, the final image having a second binning value representing the
number of pixel elements in its image pixels, wherein the first binning value is larger than the second binning value, and
wherein the calculation is based on a pixel ratio of the first binning value to the second binning value;

acquiring at least first and second final images of the sample, wherein the first final image is acquired using the calculated
exposure time, wherein the second final image is acquired using an exposure time shorter than the calculated exposure time;

identifying pixels in the first final image that are saturated;
measuring the intensity of luminescence of corresponding features in both the first and second final images, wherein measuring
provides an intensity ratio that reflects the intensity of features in the first final image in relation to the intensity
of the corresponding features in the second final image;

substituting, for any saturated pixels in the first final image, corresponding pixels from the second final image; and
adjusting the intensity of the corresponding pixels that are substituted for saturated pixels according to the intensity ratio.

US Pat. No. 9,810,703

CALIBRATION PROCESS AND SYSTEM

Bio-Rad Laboratories, Inc...

1. A package of calibration material for calibrating a medical testing machine, the package of calibration material comprising:
two different materials, at least one of which is a calibration material; a respective container for each of the materials,
the containers being connected together into a monolithic unit and maintaining separation between the materials before use
of the calibration package; and a machine-readable indicator on the outside of the package, wherein the machine-readable indicator
indicates that the package is for calibration of the medical testing machine and wherein the machine-readable indicator is
configured for automatically initiating testing of the calibration material in the package; wherein one of the materials is
a fluid configured for preparing a column of a liquid chromatography system for testing.

US Pat. No. 9,658,195

ELECTRONIC CONTROL OF PH AND IONIC STRENGTH

Bio-Rad Laboratories, Inc...

1. An apparatus for controlling pH and/or ionic strength in a vessel, the apparatus comprising,
a. a vessel in fluid communication with a first side chamber and a second side chamber, wherein the vessel does not comprise
an electrode; and

b. an anion and proton injector comprising:
the first side chamber divided from the vessel by an anion selective membrane but not by a cation selective membrane, wherein
the first side chamber comprises a first cathode; and

the second side chamber divided from the vessel by a bipolar membrane, wherein the second side chamber comprises a first anode;
wherein the first anode and first cathode form a circuit connected via a solution, when present, in the vessel and the first
and second side chambers.

US Pat. No. 9,592,483

FLUID MIXING AND RINSING SYSTEM FOR A FLOW CYTOMETER

Bio-Rad Laboratories, Inc...

1. A system for mixing deionized water and sheath fluid concentrate comprising:
a first container to contain deionized water;
a second container to contain concentrated sheath fluid having a first concentration, wherein the first container and the
second container are configured to simultaneously contain different fluids;

a pressurized reservoir;
a valve that has a first input, a second input, and an output, wherein:
the first input is fluidically coupled to the first container,
the second input is fluidically coupled to the second container,
the valve is configured to allow deionized water in the first container to flow through the output to the pressurized reservoir
when in a first position, and

the valve is configured to allow concentrated sheath fluid in the second container to flow through the output to the pressurized
reservoir when in a second position;

a pump that is fluidically interposed between the valve and the pressurized reservoir and configured to flow deionized water
in the first container and concentrated sheath fluid in the second container into the pressurized reservoir at a flowrate
that is sufficiently slow that substantially no bubbles form in the pressurized reservoir; and

a controller that is configured to maintain a concentration of a mixture of the deionized water and the concentrated sheath
fluid in the pressurized reservoir such that the mixture has a concentration of sheath fluid less than the first concentration
by repeatedly switching the valve between the first position, thereby supplying a first amount of deionized water by the pump
to the pressurized reservoir, and the second position, thereby supplying a second amount of concentrated sheath fluid by the
pump to the pressurized reservoir.

US Pat. No. 9,150,909

DETERMINATION OF THE INTEGRITY OF RNA

Bio-Rad Laboratories, Inc...

1. A method of improving accuracy and reproducibility of determining a level of integrity of a sample of ribonucleic acid
(RNA) and/or deoxyribonucleic acid (DNA) molecules, the method comprising:
receiving a first size profile of the sample of the RNA and/or DNA molecules, wherein a size profile provides a measure of
a distribution of values of at least one dimension of the RNA and/or DNA molecules, in the sample, the at least one dimension
including length and/or weight;

comparing, with an electrophoresis system, the first size profile to a plurality of reference size profiles, wherein each
reference size profile correlates to a different level of integrity; and

based on a similarity of the first size profile to one or more of the reference size profiles, determining, with the electrophoresis
system, the level of integrity of the sample of RNA and/or DNA molecules,

wherein the plurality of reference size profiles is obtained by:
at each of a plurality of different elapsed times relative to an initial time:
measuring a respective reference size profile of a reference sample of RNA and/or DNA molecules selected from a group of reference
samples having about the same initial integrity, wherein each respective reference size profile corresponds to a different
amount of degradation of the reference sample, wherein each reference size profile provides a measure of a distribution of
values of the at least one dimension of the RNA and/or DNA molecules in the reference sample; and

mapping each reference size profile to a level of integrity, a highest integrity level of reference size profile being measured
at the initial time, and each successive lower integrity level of reference size profile being measured at a longer elapsed
time from the initial time.

US Pat. No. 9,889,394

ULTRASONICALLY CLEANED LOW-PRESSURE FILTER

Bio-Rad Laboratories, Inc...

1. A method comprising:
flowing a sample through a filter device to filter the sample, wherein particulates are captured by a filter material of a
filter assembly, wherein the filter device comprises a sonotrode having a plurality of piezoelectric discs attached to the
filter assembly;

vibrating the filter device at or near a resonate frequency of the filter device to remove the particulates from the filter
material, wherein the sonotrode is operated at voltages of 5-50 Vrms and at an impedance of 20 Ohms to 500 Ohms to vibrate
the filter assembly; and

vibrating the filter device with the sonotrode in excess of 40,000 cycles without replacing the filter material,
wherein vibrating the filter device comprises monitoring the resonant or near-resonant frequency and altering power input
to the filter device with a controller to monitor the number of sonotrode cycles and to maintain a phase difference between
voltage applied to the sonotrode and current applied to the sonotrode of less than 30 degrees.

US Pat. No. 9,850,515

AFFINITY-BASED PARTITION ASSAY FOR DETECTION OF TARGET MOLECULES

Bio-Rad Laboratories, Inc...

1. A method of detecting a target molecule in a sample, the method comprising:
incubating said sample in a mixture with at least a first probe linked to a first label and a second probe linked to a second
label, wherein the first and second probes specifically bind the same target molecule, if present;

partitioning the mixture into two or more partitions, wherein on average the partitions have fewer than two probes; and
detecting the presence of the first label and the second label in at least one same partition, thereby detecting the target
molecule in the sample.

US Pat. No. 9,817,016

LOW COST OPTICAL HIGH SPEED DISCRETE MEASUREMENT SYSTEM

Bio-Rad Laboratories, Inc...

1. A system for determining an inflation rate of a droplet passing through a microfluidic channel, the system comprising:
a multicolor light source configured to emit light in two or more different wavelength bands, as temporally separated flashes,
wherein each flash comprises light of only one of said wavelength bands;

a detector configured to detect and distinguish between light in said wavelength bands, and further configured to collect
at least a first image and a second image of a droplet passing through the microfluidic channel at said different wavelength
bands; and

the microfluidic channel disposed in a translucent material,
wherein the multicolor light source, the detector, and the microfluidic channel are positioned such that light emitted by
the multicolor light source is incident upon the microfluidic channel, and such that a droplet passing through the microfluidic
channel and illuminated by the multicolor light source is imaged on the detector, and

wherein the system is configured to calculate a change in size of the droplet between the first image and the second image
by separating the first image from the second image in a video frame or a photographic exposure and determining a first diameter
of the droplet in the first image and a second diameter of the droplet in the second image.

US Pat. No. 9,815,695

IN SITU RESTORATION OF APATITE-BASED CHROMATOGRAPHY RESINS

Bio-Rad Laboratories, Inc...

1. A method for increasing the resin mass and particle strength of an apatite-based chromatography resin after purification
of a target molecule with said apatite-based chromatography resin, wherein said apatite-based chromatography resin is ceramic
hydroxyapatite (CHT), said method comprising:
(a) equilibrating said apatite-based chromatography resin;
(b) after equilibrating said apatite-based chromatography resin, loading a sample containing a target analyte onto said apatite-based
chromatography resin;

(c) after loading said sample, eluting said target analyte from said apatite-based chromatography resin by passing a solution
of elution buffer through said apatite-based chromatography resin; and

(d) after eluting said target molecule, and before subsequent loading of additional target molecule:
(i) passing a solution of calcium ion through said apatite-based chromatography resin;
(ii) after (i), passing a solution of phosphate ion having a pH of at least about 6.5 through said apatite-based chromatography
resin; and

(iii) after (ii), passing a solution of hydroxide ion through said apatite- based chromatography resin;
wherein (i), (ii), and (iii) are performed at concentrations of said calcium ions, phosphate ions, and hydroxide ions and
at volumes of said solution of calcium ion, solution of phosphate ion, and solution of hydroxide ion that are effective to
increase the resin mass and particle strength of said apatite-based chromatography resin.

US Pat. No. 9,752,177

FILTERING SMALL NUCLEIC ACIDS USING PERMEABILIZED CELLS

Bio-Rad Laboratories, Inc...

1. A method of separating DNA accessible to a DNA cleaving agent in a permeabilized cell from DNA inaccessible to the DNA
cleaving agent, the method comprising:
a) permeabilizing a cell having genomic DNA, thereby generating a permeabilized cell; introducing a DNA cleaving agent into
the permeabilized cell having genomic DNA under conditions such that the DNA cleaving agent cleaves the genomic DNA in the
cell, thereby generating cleaved DNA;

b) separating cleaved DNA that diffuses out of the intact permeabilized cell from the cell; and
c) introducing a DNA modifying agent into the permeabilized cell such that the DNA modifying agent modifies the genomic DNA
in the cell, wherein the DNA modifying agent is a DNA methyltransferase.

US Pat. No. 9,664,646

POLYACRYLAMIDE ELECTROPHORESIS GELS WITH PROTECTION AGAINST OXYGEN EXPOSURE

Bio-Rad Laboratories, Inc...

1. A combination of a polyacrylamide gel and a plastic cassette in which said polyacrylamide gel resides, the plastic cassette
comprising:
cassette walls enclosing a gel cavity and comprising a matrix plastic that is permeable to oxygen, and
a material comprising an oxygen barrier layer covering walls of said cassette such that the oxygen barrier layer is in contact
with said polyacrylamide gel, wherein the walls of the cassette are formed by injection molding and the oxygen barrier layer
is applied to the walls of the cassette by in-mold decoration,

wherein the material inhibits oxygen access to the polyacrylamide gel through the cassette walls.
US Pat. No. 9,598,725

EMULSION CHEMISTRY FOR ENCAPSULATED DROPLETS

Bio-Rad Laboratories, Inc...

1. A method of amplifying target nucleic acids in skin-encapsulated capsules comprising:
a) providing an aqueous phase comprising at least about 0.01% by weight of a skin-forming protein, an amplification reagent
mix and target nucleic acids;

b) forming an emulsion comprising droplets of said aqueous phase disposed in a continuous phase comprising an oil and a negatively
charged ionic surfactant;

c) heating said emulsion to at least about 50° C. to create an interfacial skin between each droplet and the continuous phase,
such that said droplets form skin-encapsulated capsules;

d) amplifying said target nucleic acids in said capsules; and
e) detecting amplification of said target nucleic acids.
US Pat. No. 9,164,058

POLYACRYLAMIDE GELS FOR RAPID CASTING, BLOTTING, AND IMAGING, WITH STORAGE STABILITY

Bio-Rad Laboratories, Inc...

1. A method of making a polyacrylamide gel, the method comprising:
providing a first solution comprising acrylamide/bis-acrylamide and a second solution comprising triethanolamine, an ampholyte
ampholyte selected from the group consisting of glycine and tricine, and a conjugate ampholyte consisting of an amino acid
with a pKa within the range of 8.3 to 9.6, wherein the first solution and second solution are stored separately for at least
7 days, then

forming a mixture of the first solution and the second solution and a polymerization catalyst;
adding said mixture into the mold; and
incubating the mixture in the mold under conditions sufficient to generate the polyacrylamide gel.

US Pat. No. 9,046,454

PROCESSING OF ANALYTE SUPPORTS WITH OSCILLATING FLUID BY INTERRUPTED ROTATION

Bio-Rad Laboratories, Inc...

1. A device for treating a solid member supporting an array of analytes with a process fluid, said device comprising:
a circular plate and a circular lid, said plate and lid when joined forming one or more enclosure with a cavity therein, said
cavity containing a solid member, said solid member being a blotting membrane,

an inlet port for introducing fluid into said cavity,
a support configured to rotate said plate and said lid about an axis perpendicular to said plate, said axis being centered
in said plate and lid, and

an outlet port on an outer edge of said plate relative to said axis, said outlet port for removing fluid from said cavity.
US Pat. No. 10,072,285

RNA AMPLIFICATION METHODS

Bio-Rad Laboratories, Inc...

1. A method of generating cDNA from non-polyA tailed RNA, the method comprisingadding random poly-W (A/T) or poly-S(G/C) nucleotide sequences to the 3? end of target non-polyA tailed RNA with a terminal transferase to form RNA molecules comprising degenerate nucleotide sequences at the 3? end of the molecules;
submitting the RNA molecules comprising the random poly-W or poly-S nucleotide sequences to conditions such that the RNA molecules comprising the random poly-W or poly-S nucleotide sequences anneal to form a duplex of a first and second RNA molecule annealed to each other via the random poly-W or poly-S nucleotide sequences; and
extending the 3? ends of the molecules in the duplex with a DNA polymerase such that the 3? ends act as a primer to form (i) a first cDNA complementary to the first RNA molecule of the duplex and (ii) a second cDNA complementary to the second RNA molecule of the duplex, wherein the first and second cDNAs further comprise a 5? sequence that is a complement of the random poly-W or poly-S nucleotide sequence.

US Pat. No. 9,885,862

ADJUSTABLE DIGITAL MICROSCOPE DISPLAY

Bio-Rad Laboratories, Inc...

1. An adjustable imaging apparatus comprising:
a sample stage;
an elevated assembly positioned above the sample stage, configured to move along at least one range of motion relative to
the sample stage and to be adjustable along at least two degrees of freedom, wherein a first degree of freedom is a first
turning axis along a first edge of the elevated assembly and wherein a second degree of freedom is a second turning axis along
a second edge of the elevated assembly parallel to the first turning axis of the elevated assembly;

one or more light shields, mechanically coupled to the elevated assembly, the one or more light shields being configured to
move between a first extended shielding position that extends from the elevated assembly and a second extended shielding position
that extends from the elevated assembly, so as to block ambient light from being incident on the sample stage; and

an imaging camera apparatus, positioned within the sample stage to capture images of objects on the sample stage.

US Pat. No. 9,689,840

CONJUGATES OF 1,4,7-TRIAZACYCLONONANES, DINUCLEAR METAL COMPLEXES OF SUCH CONJUGATES, AND METHODS OF USE FOR BOTH 1,4,7-TRIAZACYCLONONANES AND CONJUGATE

Bio-Rad Laboratories, Inc...

1. A method for the preparation of a separation medium for use in electrophoresis of phosphorylated species, said method comprising
polymerizing a monomer mixture comprising acrylamide, a crosslinker, an initiator, and a compound having the formula

in which:
one of R1 through R6 is -L-R7 in which L is a member selected from the group consisting of —CH2—C(?O)—NH—(C1-C4 alkyl)—, —CH2—C(?O)—NH—(C1-C4 alkyl)—NH—, and —(C1-C4 alkyl)—NH—, and R7 is an acrylamide group;

the remainder of R1 through R6 are independently selected from the group consisting of H and C1-C6 alkyl; and

M is a divalent metal.

US Pat. No. 9,687,848

MICROFLUIDIC SYSTEM WITH FLUID PICKUPS

Bio-Rad Laboratories, Inc...

11. A method of processing fluid, the method comprising:
selecting a microfluidic device including at least one well component, the at least one well component providing an input
well and an output well, the microfluidic device also including a channel component attached directly to a top side of the
at least one well component, the channel component including

(a) a body having a top surface, wherein a plurality of microchannels are formed in the top surface, and wherein the plurality
of microchannels meet one another at a channel intersection,

(b) a cover bonded to the top surface of the body and forming a top wall of each of the plurality of microchannels,
(c) an input tube projecting from a bottom surface of the body and located in the input well, and
(d) a sample port that communicates with the input well separately from the input tube;
introducing, with a fluid-transfer device, a sample-containing fluid into the input well through the sample port and into
contact with a bottom end of the input tube;

operatively connecting at least one vacuum/pressure source to the microfluidic device; and
creating a pressure differential with the at least one vacuum/pressure source that drives at least a portion of the sample-containing
fluid from the input well to the output well on a path defined by the channel component, wherein the path extends upward through
the input tube, through the channel intersection, and downward to the output well.

US Pat. No. 9,527,049

STABILIZED DROPLETS FOR CALIBRATION AND TESTING

Bio-Rad Laboratories, Inc...

1. A method comprising the steps of:
(a) combining a first droplet population with a second droplet population to form a combined droplet population;
(b) mixing the combined droplet population in a mixing module to produce a mixture in which each of the first droplet population
and the second droplet population remains substantially intact;

(c) stabilizing droplets of the first droplet population and droplets of the second droplet population by heating that forms
a droplet skin including a skin-forming protein; and

(d) shipping at least a portion of the combined droplet population at least one kilometer after the steps of mixing and stabilizing.
US Pat. No. 9,506,939

STABILIZATION OF LABILE ANALYTES IN REFERENCE MATERIALS

Bio-Rad Laboratories, Inc...

1. A multianalyte control kit comprising
a first container holding at least one unstable control analyte, wherein the at least one unstable control analyte is lyophilized
in the form of beads; and

a second container holding at least one stable control analyte in a base matrix solution from a biological sample from which
the unstable control analyte has been removed, wherein the biological sample is saliva, lymph, urine, milk, mucus, cerebrospinal
fluid (CSF), cell lysate, or tissue culture supernatant, wherein the base matrix solution comprises a buffer selected from
HEPES and Tris, wherein the buffer is in an amount to control the pH of the base matrix solution in a range from 6 to 8.

US Pat. No. 10,107,780

DIP-STICK WESTERN BLOT

Bio-Rad Laboratories, Inc...

1. A method of separating and detecting analytes of a sample, the method comprising:applying a sample to a separation medium, wherein the separation medium is a polyacrylamide gel;
separating analytes of the sample in the separation medium to generate a distribution of the analytes along a separation axis;
immobilizing the analytes on a dipstick embedded in part of the separation medium in which analytes become distributed such that the dipstick or an elongated portion thereof is aligned parallel to the separation axis, wherein distribution of analytes immobilized on the dipstick preserves the distribution of analytes as occurred in the separation medium, wherein the dipstick is embedded before the separating;
removing the dipstick from the separation medium; and
detecting the analytes immobilized on the removed dipstick,
wherein separating analytes of the sample comprises performing electrophoresis, electroosmosis, or isoelectric focusing.
US Pat. No. 9,994,887

METHODS AND COMPOSITIONS FOR IMPROVING EFFICIENCY OF NUCLEIC ACIDS AMPLIFICATION REACTIONS

Bio-Rad Laboratories, Inc...

1. A method of amplifying a target nucleic acid in a sample, the method comprising,incubating said target nucleic acid in a reaction mixture comprising:
at least one primer,
an exonuclease deficient hybrid polymerase having portions of two parental polymerases, the exonuclease deficient hybrid polymerase comprising a polymerase domain and a heterologous DNA binding domain, wherein the polymerase domain is from a B family polymerase, and
a double-stranded DNA binding dye,
wherein said incubating is under conditions that permit amplification of said target nucleic acid by said hybrid polymerase; and
using quantitative polymerase chain reaction (qPCR) detection to monitor amplification of the target nucleic acid, wherein signal from the double-stranded DNA binding dye is detected in real-time;
thereby amplifying a target nucleic acid in a sample.

US Pat. No. 9,921,193

BOTTLE PRESSURIZATION DELIVERY SYSTEM

Bio-Rad Laboratories, Inc...

1. A container assembly for holding a fluid medium, the container assembly comprising:
an external container having a first port and a second port, the external container defining an interstitial volume fluidly
connected to the first port, wherein the external container is formed of a semi-rigid material having walls configured to
restorably flex, expanding when under pressure and returning to an unexpanded state when not under pressure;

an internal container contained within the external container and being fluidly sealed from the interstitial volume, the internal
container holding a fluid medium and being fluidly connected to the second port by way of a valve, wherein the internal container
is formed of a flexible material;

a sensor mechanically coupled to the external container, configured to determine a volume of the fluid medium held within
the internal container; and

a non-transitory computer-readable memory, mechanically coupled to the external container, that stores updatable information
regarding the fluid medium held in the container assembly.

US Pat. No. 9,885,034

METHODS AND COMPOSITIONS FOR NUCLEIC ACID ANALYSIS

Bio-Rad Laboratories, Inc...

1. A method for sample preparation, comprising:
a. combining:
i. a first droplet comprising an oligonucleotide barcode and
ii. a target analyte
into a partition, wherein said first droplet is burstable, fusible, or mergeable upon application of a treatment; and
b. applying said treatment to said first droplet to release said oligonucleotide barcode to said target analyte.
US Pat. No. 9,809,851

SYSTEMS AND METHODS FOR SEQUENCING IN EMULSION BASED MICROFLUIDICS

Bio-Rad Laboratories, Inc...

1. A method of identifying primer/target nucleic acid partitions as being generated from a same target nucleic acid partition
mixture drop, the method comprising:
providing a plurality of mixture drops, each mixture drop including copies of at least one target nucleic acid;
generating a plurality of reaction partitions from each mixture drop, the reaction partition including one or more primers,
wherein one or more marker reagent is present in each reaction partition such that each primer set in a reaction partition
is represented by a pre-determined and known unique signal based on the one or more marker reagent identity and/or concentration;
and

detecting signal from the droplets with a detector, wherein the signal includes a signal whether at least one primer in a
partition hybridizes to a target nucleic acid;

receiving in a computer a data signal obtained by the detector over a time period, the data signal including signals from
a plurality of reaction partitions generated from a plurality of mixture drops, each mixture drop corresponding to a cohort
of partitions and including copies of at least one target nucleic acid, each partition including one or more primers, wherein
the data signal includes data about whether at least one primer in a partition hybridizes to a target nucleic acid;

identifying a hybridization status of each partition based on the respective signal of the respective partition, the hybridization
status of a partition indicating whether a primer in the partition hybridized to a target nucleic acid, wherein a signal of
a particular partition corresponds to a particular time;

for each of a plurality of particular times in the time period:
for a time window around the particular time:
calculating with the computer an amount of partitions that have contradictory hybridization statuses and include a same primer,
thereby obtaining a temporal function;

identifying extrema in the temporal function; and
determining a cohort of successive partitions occurring between corresponding extrema in the temporal function as corresponding
to a same mixture drop.

US Pat. No. 9,791,362

FLUID MIXING AND RINSING SYSTEM FOR A FLOW CYTOMETER

Bio-Rad Laboratories, Inc...

1. A method of mixing deionized water and concentrated sheath fluid in a flow cytometer comprising:
supplying deionized water from a first container;
supplying concentrated sheath fluid from a second container;
pumping the deionized water and the concentrated sheath fluid into a reservoir at a rate that is sufficiently slow to substantially
eliminate turbulence that causes bubbles to form in the reservoir; and

controlling a first valve that is connected to the first container in a first position and the second container in a second
position so that the first valve is disposed in the first position until a predetermined amount of the deionized water is
supplied to the reservoir and the first valve is disposed in the second position until a predetermined amount of the concentrated
sheath fluid is supplied to the reservoir to produce sheath fluid having a predetermined concentration in the reservoir.

US Pat. No. 9,776,183

MICROFLUIDIC CARTRIDGE DEVICES AND METHODS OF USE AND ASSEMBLY

Bio-Rad Laboratories, Inc...

1. A microfluidic cartridge interface device comprising:
an enclosure configured to receive a microfluidic cartridge within a receptacle thereof, the microfluidic cartridge having
one or more pressure septa in fluid communication with a network of channels to facilitate controlled flow of one or more
reagents and/or a fluid sample through the network of channels upon introduction of pressure through the one or more pressure
septa;

an alignment feature within the receptacle configured to engage with a corresponding alignment feature of the microfluidic
cartridge to facilitate alignment of the microfluidic cartridge within the receptacle; and

one or more pressure needles arranged at pre-determined positions corresponding to the one or more pressure septa of the microfluidic
cartridge when aligned within the receptacle.

US Pat. No. 9,774,804

DIGITAL IMAGING WITH MASKED PIXELS

Bio-Rad Laboratories, Inc...

1. An imaging device comprising:
an image sensor comprising an array of pixels;
a processor coupled with the image sensor; and
a mask coupled with the image sensor, the mask configured to darken a plurality of isolated pixels or groups of pixels interspersed
within the array of pixels;

wherein the array of pixels comprises a plurality of un-darkened pixels, and the processor is configured to:
receive image data from the image sensor including a signal for each pixel in the array of pixels, and determine a dark current
fixed pattern noise based on the image data received from the plurality of darkened pixels or groups of pixels;

for each of the plurality of un-darkened pixels in the array of pixels, determine nearby darkened pixels that are nearest
to each of the plurality of un-darkened pixels;

determine a change in a dark current fixed pattern of each of the nearby darkened pixels from a previously recorded dark current
fixed pattern of each of the nearby darkened pixels;

determine a change in a dark current fixed pattern for each of the plurality of un-darkened pixels from a previously recorded
dark current fixed pattern of each of the plurality of un-darkened pixels, based on an interpolation of the change in the
dark current fixed pattern of each of the nearby darkened pixels that are nearest to each of the plurality of un-darkened
pixels;

determine a dark current fixed pattern of each of the plurality of un-darkened pixels based on the change in a dark current
fixed pattern for each of the plurality of un-darkened pixels; and

subtract the dark current fixed pattern of each of the plurality of un-darkened pixels from the signal for each of the plurality
of un-darkened pixels.

US Pat. No. 9,669,402

ANIONIC EXCHANGE-HYDROPHOBIC MIXED MODE

Bio-Rad Laboratories, Inc...

1. A method of purifying a biomolecule from a sample, the method comprising
contacting the sample to a chromatographic solid support linked to a ligand according to formula (I):
Support-(X)—N(R1,R2)—R3-L-Ar, or a salt thereof  (I),
wherein
X is a spacer or absent;
R1 and R2 are each selected from hydrogen and an alkyl comprising 1-6 carbons;

R3 is an alkyl comprising 1-6 carbons or a cyclo alkyl comprising 1-6 carbons;

L is NR4, O, or S; wherein R4 is hydrogen or an alkyl comprising 1-6 carbons;

Ar is an aryl; and
the nitrogen in the N(R1, R2) is positively charged during the contacting; and

collecting a purified biomolecule.
US Pat. No. 9,594,054

TARGETED DELIVERY OF REAGENTS TO SPOTS ON A PLANAR SUPPORT THROUGH PATTERNED TRANSFER SHEETS

Bio-Rad Laboratories, Inc...

1. A method for determining whether an antigen is immobilized on a blotting membrane by contacting an antibody with said antigen,
said method comprising:
(a) placing said blotting membrane in contact with a porous hydrophilic sheet having a laterally delimited region therein
that is bordered by a hydrophobic or impermeable barrier and in which said antibody is retained, while aligning said laterally
delimited region with a spot on said blotting membrane where said antigen resides if so immobilized on said blotting membrane;

(b) causing transfer of said antibody from said laterally delimited region of said porous hydrophilic sheet to said spot,
wherein the causing transfer comprises placing a first buffer solution-wetted sheet of absorbent material on a side of said
blotting membrane opposite the side in contact with said porous hydrophilic sheet, and a second buffer solution-wetted sheet
of absorbent material on a side of said porous hydrophilic sheet opposite the side in contact with said blotting membrane,
to form a transfer stack, and passing an electrical current across said transfer stack in a direction causing said transfer;
and

(c) detecting any occurrence of a binding reaction between said antibody and said antigen as an indication of the presence
of said antigen in said spot.

US Pat. No. 9,551,637

FLOW RATE BALANCED, DYNAMICALLY ADJUSTABLE SHEATH DELIVERY SYSTEM FOR FLOW CYTOMETRY

Bio-Rad Laboratories, Inc...

1. A method of controlling pressure of a sheath fluid in a positively pressurized reservoir holding a volume of sheath fluid
and a volume of positively pressurized air in a flow cytometer configured to flow sheath fluid from the positively pressurized
reservoir through a nozzle at an out-flow rate, the method comprising:
continuously pumping, using a pump interposed between an external container and the positively pressurized reservoir, the
sheath fluid from the external container into the positively pressurized reservoir to maintain a substantially constant sheath
fluid level in the positively pressurized reservoir so that an in-flow rate of sheath fluid flowing into the positively pressurized
reservoir is substantially equal to the out-flow rate of the sheath fluid flowing out of the nozzle;

determining the out-flow rate of fluid flowing out of the nozzle based on data from a visual sensor configured to detect droplet
locations of the sheath fluid flowing out of the nozzle;

controlling the pressure of the volume of positively pressurized air based upon the determination, so that the out-flow rate
of fluid flowing out of the nozzle, as determined by the visual sensor, remains substantially constant; and

adjusting the in-flow rate of the sheath fluid flowing into the positively pressurized reservoir by adjusting a pump speed
of the pump whenever the substantially constant sheath fluid level changes.

US Pat. No. 9,488,616

GELC-MS USING STAIN FREE TECHNOLOGY

Bio-Rad Laboratories, Inc...

1. A method of preparing a protein sample for mass spectroscopy, wherein said protein sample is a complex protein sample comprising
at least 50 different proteins, said method comprising:
a) providing a one-dimensional (1D) electrophoresis gel comprising said complex protein sample, wherein proteins of said complex
protein sample have been separated by electrophoresis in only one direction, and at least some proteins comprise tryptophan
residues;

b) contacting said complex protein sample of step a) with a halo-substituted compound;
c) exposing said 1D electrophoresis gel to UV light, thereby reacting said tryptophan residues with said halo-substituted
compound, covalently modifying said tryptophan residues, and rendering said tryptophan residues fluorescent;

d) detecting fluorescence emitted from said 1D electrophoresis gel of step c);
e) excising at least one portion of said 1D electrophoresis gel based upon detected fluorescence of step d), wherein said
at least one portion comprises proteins of said complex protein sample and is excised from one lane of said 1D electrophoresis
gel, wherein said excising comprises making cuts in said 1D electrophoresis gel around said at least one portion, thereby
separating said at least one portion from the rest of said 1D electrophoresis gel; and

f) subjecting proteins from said at least one portion of said 1D electrophoresis gel of step e) to mass spectroscopy, wherein
more proteins are subjected to mass spectroscopy and detected by mass spectroscopy than would be if said 1D electrophoresis
gel were contacted with a protein stain.

US Pat. No. 9,329,127

FLUORESCENCE SCANNING HEAD WITH MULTIBAND DETECTION

Bio-Rad Laboratories, Inc...

1. A scanning system for detection and discrimination of a plurality of targets in each of a plurality of samples, each target
bearing a distinct fluorescent label, said scanning system comprising:
a sample plate with a planar array of sample wells disposed therein;
a scanning head comprising:
(i) a light source actuatable to supply excitation light at a plurality of excitation wavelengths, each of said excitation
wavelengths selected to excite one of said fluorescent labels;

(ii) a plurality of multiband emission filters, each arranged to receive emission light emitted by two or more of said fluorescent
labels upon excitation of said labels by said excitation light, and to pass said emission light so received in individually
distinguishable wavelength bands, each said wavelength band including emission light from one of said fluorescent labels;
and

(iii) a separate detector for each said multiband emission filter, wherein the detector is a non-imaging sensor that converts
light impinging thereon to a measurable signal, said detector arranged to receive emission light passing through said multiband
emission filter; and

drive motors or mechanisms for causing translational movement of said scanning head over each of said sample wells in succession,
wherein different multiband emission filters pass light in different combinations of wavelength bands, the combinations do
not duplicate wavelength bands, said plurality of said multiband emission filters collectively passes light emitted by all
of said fluorescent labels.

US Pat. No. 10,101,296

MINI-GEL COMB

Bio-Rad Laboratories, Inc...

1. A mold for casting and retaining electrophoresis gel strips, the mold comprising:an array of elongated lane cavities, each having a length greater than a width, each lane cavity having a top end, a bottom end, and two or more sides disposed along the length of the lane cavities, the lane cavities oriented parallel to each other;
a matrix comprising at least one solid material and disposed along the sides of the lane cavities, the matrix separating the lane cavities from each other and from a space outside the mold; and
a plurality of windows, wherein two windows are associated with each lane cavity and are disposed on opposite sides of the lane cavity orthogonal to the length of the lane cavities, each window running lengthwise along the lane cavity and penetrating the matrix, wherein:
the windows associated with each lane cavity form passages from the lane cavity through the matrix to the space outside the mold,
the matrix comprises a first solid material sandwiched between two pieces of a second solid material, the first solid material separating the lane cavities from each other, and the two pieces of the second solid material at least partially separating the lane cavities from the space outside the mold,
the first solid material comprises a comb, the comb comprising a plurality of teeth, and the lane cavities comprise the spaces between the teeth, and
the second solid material comprises two window plates, the window plates bonded to opposite sides of the comb, and each window plate comprising at least one of the plurality of windows,
thereby forming an opening from each lane cavity through the matrix to an adjacent lane cavity or the space outside the mold.
US Pat. No. 9,802,822

APATITE PRETREATMENT

Bio-Rad Laboratories, Inc...

1. A method of treating an apatite solid surface prior to first use in a chromatographic procedure, the method comprising:
(a) providing an apatite solid surface, wherein the apatite surface has not previously been used in any chromatographic procedure
and has not previously been contacted with any sample;

(b) contacting the apatite solid surface with a phosphate buffered solution at a pH of at least about 6.5; and then
(c) contacting the apatite solid surface obtained in step (b) with a solution having an alkaline metal hydroxide: and then
(d) purifying protein from a sample with the apatite solid surface obtained in step (c).

US Pat. No. 9,794,454

CONTACT IMAGER

Bio-Rad Laboratories, Inc...

1. An imaging cassette comprising a base support, an image sensor, and a light-tight lid, wherein the imaging cassette is
configured to receive a piece of X-ray film and sample membrane when the image sensor is present in the imaging cassette,
wherein the imaging cassette is configured to receive the sample membrane between the X-ray film and the image sensor or wherein
the imaging cassette is configured to receive the sample membrane between the X-ray film and the light-tight lid.

US Pat. No. 9,784,664

MULTIDIMENSIONAL HYDRODYNAMIC FOCUSING CHAMBER

Bio-Rad Laboratories, Inc...

1. A method of hydrodynamically focusing a fluid sample containing one or more particles, the method comprising:
streaming a sheath fluid through a first microfluidic channel;
passing the sheath fluid into an inline chamber having a central island projecting into the chamber, wherein the island has
smaller lateral and vertical dimensions than the chamber, thereby leaving a gap at each lateral side of the island to form
side paths, and a gap above a top surface of the island to form an overhead path, such that the sheath fluid flows around
the island on the side paths and over the island on the overhead path;

introducing a particle-containing fluid sample into the chamber through an aperture in the island, such that the fluid sample
is carried downstream along the top surface of the island with the sheath fluid and confined radially by the sheath fluid;
and

passing the sheath fluid and fluid sample into a second microfluidic channel.

US Pat. No. 9,766,207

AFFINITY METHODS AND COMPOSITIONS EMPLOYING ELECTRONIC CONTROL OF PH

Bio-Rad Laboratories, Inc...

1. A method for selectively positioning a target analyte in a solution containing a mixture of analytes, the target analyte
having a pH dependent charge, the method comprising:
providing into a chamber a sample comprising the solution containing the mixture of analytes, including the target analyte,
wherein the chamber comprises a first and a second electrode and at least two proton/hydroxyl injectors positioned between
the electrodes;

generating a first pH step with one of the at least two proton/hydroxyl injectors, thereby generating a first sub-area having
the first pH step, and applying an electric field across the electrodes, thereby moving a portion of the mixture of analytes
to the first sub-area in the chamber based on the pH dependent charge of the target analyte, wherein the portion of the mixture
of analytes comprises analytes having a range of isoelectric points from of about pH 3 or more to pH of about 10 or less;
and

generating a second pH step with the second proton/hydroxyl injector, thereby generating a second sub-area having the second
pH step, wherein the second pH step is narrower than the first pH step and encompasses a pH range that includes the pI of
the target analyte, thereby selectively positioning the target analyte in a sub-area near the second proton/hydroxyl injector
of the chamber.

US Pat. No. 9,663,766

METHODS FOR PURIFYING ADENOVIRUS VECTORS

Bio-Rad Laboratories, Inc...

1. A method of purifying an adenovirus from an impure preparation, the method comprising:
(a) contacting an impure preparation comprising the adenovirus to a mixed mode chromatography support, wherein the mixed mode
chromatography support is a hydrophobic cation exchange chromatography support, under conditions that allow the adenovirus
to bind to the mixed mode chromatography support;

(b) eluting the adenovirus from the mixed mode chromatography support, thereby forming a mixed mode eluate;
(c) contacting the mixed mode eluate to an anion exchange chromatography support under conditions that allow the adenovirus
to bind to the anion exchange support; and

(d) eluting the adenovirus from the anion exchange chromatography support.
US Pat. No. 9,623,414

LOCALIZED TEMPERATURE CONTROL FOR SPATIAL ARRAYS OF REACTION MEDIA

Bio-Rad Laboratories, Inc...

1. Apparatus for performing polymerase chain reactions in a plurality of samples, said apparatus comprising:
a plurality of thermally conductive sample blocks for polymerase chain reactions, arranged in a fixed horizontal array, wherein
each sample block comprises a plurality of sample wells and is configured to retain a plurality of samples;

a plurality of independently controlled thermoelectric modules, a thermoelectric module positioned underneath each said sample
block, wherein the thermoelectric modules are configured to cycle the temperatures of the sample blocks for polymerase chain
reactions;

a layer of thermally conductive material between each sample block and each thermoelectric module; and
a solid barrier of thermally insulating material positioned between each pair of adjacent sample blocks to thermally isolate
the sample blocks of the pair from each other.

US Pat. No. 9,556,423

PCR REACTION MIXTURES WITH DECREASED NON-SPECIFIC ACTIVITY

BIO-RAD LABORATORIES, INC...

1. A reaction mixture comprising:
a polymerase conjugated to a DNA binding domain, wherein the DNA binding domain is a Sso7 domain; and
from about 5 mM to about 15 mM of free arginine, or a salt thereof.

US Pat. No. 9,513,264

ELECTRONIC CONTROL OF PH AND IONIC STRENGTH

Bio-Rad Laboratories, Inc...

1. An apparatus for controlling pH and/or ionic strength in a vessel, the apparatus comprising,
a. a vessel in fluid communication with a first side chamber and a second side chamber, wherein the vessel does not comprise
an electrode; and

b. an anion and proton injector comprising:
the first side chamber divided from the vessel by an anion selective membrane but not by a cation selective membrane, wherein
the first side chamber comprises a first cathode; and

the second side chamber divided from the vessel by a bipolar membrane, wherein the second side chamber comprises a first anode;
wherein the first anode and first cathode form a circuit connected via a solution, when present, in the vessel and the first
and second side chambers.

US Pat. No. 9,482,213

COMMON MODE PULSE DAMPER FOR RECIPROCATING PUMP SYSTEMS

Bio-Rad Laboratories, Inc...

1. A pressure vessel for use at outlets of two piston pumps, said pressure vessel comprising a first flow-through compartment
with a first inlet port and a first outlet port, and a second flow-through compartment with a second inlet port and a second
outlet port, said first and second flow-through compartments separated and sealed from each other by a diaphragm that is flexible
and impermeable to fluid, wherein first and second fluid conduits connected respectively to the first and second outlet ports
are in direct fluid communication with a common outlet in which fluids from the two piston pumps are combined.

US Pat. No. 9,841,378

SERIES ABSORBANCE GLASS AND INTERFERENCE FILTERS

Bio-Rad Laboratories, Inc...

1. An imaging assembly comprising:
a sample platform having a target region;
an excitation light module arranged proximate to the sample platform;
a lens module arranged to receive light emitted from the target region, wherein the lens module includes two or more lenses
arranged in an uninterrupted series as an optical baffle within a lens barrel, wherein the lenses in the lens module decrease
in diameter along the optical path of the emission light and are configured to refract off-normal light toward interior walls
of the lens barrel;

a first series filter assembly, positioned along an optical path in line with the lens module, positioned in front of the
lens module along the optical path, the first series filter assembly being comprised of an absorbance glass having at least
one side coated with a multi-band thin film interference filter layer;

a light sensor arranged to receive light emitted from the lens module; and
an imaging module configured to process data captured by the light sensor.
US Pat. No. 9,840,697

FUSION POLYMERASES

Bio-Rad Laboratories, Inc...

1. A polypeptide having polymerase activity and 5?-3? exonuclease activity, the polypeptide comprising a 5?-3? exonuclease
domain linked to a heterologous polymerase that does not naturally have 5?-3? exonuclease activity, wherein the heterologous
polymerase comprises a family B polymerase catalytic domain having polymerase activity and the 5?-3? exonuclease domain is
a flap endonuclease, wherein the polypeptide substantially lacks 3?-5? exonuclease activity.
US Pat. No. 9,822,393

COMPOSITIONS, METHODS AND SYSTEMS FOR POLYMERASE CHAIN REACTION ASSAYS

Bio-Rad Laboratories, Inc...

1. A method of detecting a target nucleic acid, the method comprising:
(a) providing an oil composition comprising an oil and a fluorosurfactant mixture, wherein said fluorosurfactant mixture includes
a triblock copolymer comprising polyethylene glycol (PEG) polymer covalently linked by an ester bond to polyhexafluoropropylene
oxide (PFPE) at both ends,

a diblock copolymer comprising a PEG block covalently linked by an ester bond to a PFPE block at only one end, and
a PFPE carboxylic acid;
(b) providing an aqueous composition comprising a target nucleic acid, a non-ionic non-fluorosurfactant and an intercalating
dye;

(c) contacting the oil composition of a) with the aqueous composition of b), thereby generating a plurality of droplets suspended
in a continuous phase;

(d) thermally cycling said plurality of droplets to amplify said target nucleic acid; and
(e) detecting said target nucleic acid, wherein said detecting comprises measuring fluorescence from said intercalating dye.

US Pat. No. 10,022,721

MICROFLUIDIC DEVICES AND METHODS OF THEIR USE

Bio-Rad Laboratories, Inc...

1. A system for labeling and alternating flow of differentially-labeled droplets, the system comprising:at least one inlet for droplets within a continuous phase,
at least two microfluidic channels that are connected to the at least one inlet at one end and form a junction with a joint microfluidic channel at the other end,
a connection channel configured to allow the continuous phase fluid but not the droplets to flow between the microfluidic channels, wherein the connection channel is sized such that the droplets are too large to enter one of the microfluidic channels, and wherein the connection channel is configured to equalize pressure between the two microfluidic channels, and
a labeling device configured to label droplets in one of the microfluidic channels;
wherein when the droplets flow through the at least two microfluidic channels toward the joint microfluidic channel, the labeling device labels the droplets that flow in one of the microfluidic channels.
US Pat. No. 9,944,998

GENETIC ASSAYS

Bio-Rad Laboratories, Inc...

1. A method of quantifying previously recombined nucleic acids containing a first sequence and a second sequence, the method comprising:a. obtaining a sample comprising viral genomic nucleic acids, wherein the viral genomic nucleic acids comprise (i) first parental nucleic acids representing a first viral genome that includes the first sequence and not the second sequence, (ii) second parental nucleic acids representing a second viral genome that includes the second sequence and not the first sequence, and (iii) a plurality of previously recombined nucleic acids including, on a same strand, the first sequence and the second sequence;
b. partitioning the sample into more than about 1,000 compartments such that each compartment of only a subset of the more than about 1,000 compartments contains the first sequence, each compartment of only a subset of the more than about 1,000 compartments contains the second sequence, and each compartment of only a subset of the more than about 1,000 compartments contains both the first sequence and the second sequence;
c. performing amplification of the first sequence and amplification of the second sequence within the more than about 1,000 compartments;
d. detecting at least one amplification signal from the more than about 1,000 compartments indicating whether the first sequence is present in a given compartment and whether the second sequence is present in the given compartment;
e. enumerating compartments of the more than about 1,000 compartments that comprise the first sequence but not the second sequence to obtain a first value, the second sequence but not the first sequence to obtain a second value, both the first sequence and the second sequence to obtain a third value, and neither the first sequence nor the second sequence to obtain a fourth value, wherein the step of enumerating is performed using the at least one amplification signal detected from the more than about 1,000 compartments;
f. calculating an expected number of compartments of the more than about 1,000 compartments comprising both the first sequence and the second sequence by chance co-localization, using the first, second, and fourth values; and
g. adjusting the third value of step (e) using the expected number of step (f) in order to obtain a numerical value for the recombined nucleic acids.
US Pat. No. 9,702,885

RECOMBINANT DEAMIDATED GLIADIN ANTIGEN

Bio-Rad Laboratories, Inc...

1. An antigen for detecting celiac disease, the antigen comprising: a recombinant deamidated gliadin fusion protein consisting
of SEQ ID NO:7 or SEQ ID NO:8; wherein the recombinant deamidated gliadin fusion protein is immobilized on a solid support
and is capable of binding to anti-deamidated gliadin antibodies.

US Pat. No. 9,694,360

FLUID MANIPULATOR HAVING FLEXIBLE BLISTER

Bio-Rad Laboratories, Inc...

1. A system for fluid manipulation, the system comprising:
a composite wafer having a flexible layer and a substantially rigid layer adhered to the flexible layer, the flexible layer
defining one or more recesses that are covered by the substantially rigid layer to form one or more reservoirs and one or
more fluid channels among the one or more reservoirs, wherein the substantially rigid layer also defines at least one fluid
flow channel within the substantially rigid layer; and

a movable compression device in contact with the flexible layer and configured to progressively compress the flexible layer
such that when the movable compression device traverses the flexible layer, fluid is forced through the one or more fluid
channels and the one or more reservoirs in a sequence determined by the layout of the one or more fluid channels and the one
or more reservoirs;

wherein the movable compression device is a roller, and wherein the fluid flow channel defined within the substantially rigid
layer is positioned such that the roller forces fluid through the fluid flow channel within the substantially rigid layer
past the roller in a direction counter to the direction of motion of the roller.

US Pat. No. 9,671,402

LATERAL FLOW BLOTTING ASSAY

Bio-Rad Laboratories, Inc...

1. A porous substrate having a length and a width and comprising a reagent reservoir region and a lateral flow region, said
reagent reservoir region comprising or bordered by an impermeable or hydrophobic barrier, said impermeable or hydrophobic
barrier substantially blocking flow of a liquid from the reagent reservoir region into the lateral flow region until lateral
flow is initiated, wherein
said reagent reservoir region comprises at least a first reagent reservoir sub-region and a second reagent reservoir sub-region,
wherein the first and second reagent reservoir sub-regions are not in fluid communication and are separated by a hydrophobic
or impermeable barrier that is parallel relative to flow direction; and

said lateral flow region comprises at least a first lateral flow sub-region and a second lateral flow sub-region, wherein
said first and second lateral flow sub-regions are not in fluid communication and are separated by a hydrophobic or impermeable
barrier that is parallel relative to flow direction, wherein the first reagent reservoir sub-region is configured to deliver
reagents therein to the first lateral flow sub-region and the second lateral flow sub-region is configured to deliver reagents
therein to the second reagent reservoir sub-region.

US Pat. No. 10,048,270

HCP ANTISERUM VALIDATION USING A NON-INTERFERING PROTEIN STAIN

Bio-Rad Laboratories, Inc...

1. A method for validating an immunological detection reagent comprising a polyclonal antibody mixture for use in detection of contaminating host cell proteins (HCPs) in a biological sample, the method comprising:providing a biological sample containing HCPs;
separating the HCPs by size and isoelectric point;
transferring the separated HCPs to a membrane, thereby producing membrane-bound HCPs;
staining membrane-bound HCPs using a non-interfering total protein stain, thereby detecting host cell protein (HCP) positions on the membrane with the non-interfering total protein stain;
observing and recording the HCP positions on the membrane detected by the non-interfering total protein stain;
contacting the membrane with the polyclonal antibody mixture, thereby binding antibodies from the polyclonal antibody mixture to the membrane-bound HCPs;
detecting antibodies from the polyclonal antibody mixture bound to the membrane, thereby detecting HCP positions on the membrane with the polyclonal antibody mixture;
observing and recording the HCP positions on the membrane detected by the polyclonal antibody mixture; and
comparing the HCP positions detected by the non-interfering total protein stain to the positions detected by the polyclonal antibody mixture, thereby validating the polyclonal antibody mixture for use in detection of contaminating host cell proteins HCPs in a biological sample.

US Pat. No. 9,846,111

OPTICAL DETECTION SYSTEM FOR PARTICLES

Bio-Rad Laboratories, Inc...

1. A detection system for particles, comprising:
a channel;
a light source configured to generate light;
one or more optical elements configured to focus a beam of the light on an irradiation zone within the channel;
a mask operatively disposed in an optical path between the light source and the channel, wherein the mask is configured to
block a portion of the beam, thereby producing a shadow region;

an optical slit operatively disposed in the optical path between the light source and the channel at a position optically
upstream of the mask; and

a detector configured to detect light deflected from the beam into the shadow region by interaction with a particle passing
through the irradiation zone;

wherein the mask is a line mask including a masking region elongated parallel to the optical slit and elongated orthogonal
to a flow direction in the channel through the irradiation zone.

US Pat. No. 9,822,396

CHROMOSOME CONFORMATION CAPTURE IN PARTITIONS

Bio-Rad Laboratories, Inc...

1. A method for detecting chromosomal DNA sequence elements that are in close proximity in a cell or in an intact nucleus,
the method comprising:
a) cross-linking the chromosomal DNA in the cell or the intact nucleus, thereby forming a cross-linked DNA product in which
chromosomal DNA sequence elements that are in close proximity in the cell or the intact nucleus are cross-linked;

b) obtaining the cross-linked DNA product;
c) digesting the cross-linked DNA product with an endonuclease to form a mixture containing a plurality of cross-linked DNA
digest fragments;

d) partitioning the cross-linked DNA digest fragments to create a plurality of partitions, wherein the partitions are droplets;
and

e) detecting the presence of two or more DNA sequence elements that are distal with respect to the primary sequence of the
genome in a single droplet partition, wherein detection of two or more DNA sequence elements in a single droplet partition
indicates that the two or more DNA sequence elements were in close proximity in the cell or intact nucleus.

US Pat. No. 9,804,149

PATIENT-BASED RESULTS DISPLAY

Bio-Rad Laboratories, Inc...

1. A medical testing machine, comprising:
an input location, an output location, and a vial handling mechanism that moves vials of media sampled from patients from
the input location through the testing machine to the output location;

an extraction mechanism that extracts the media from each vial in turn for testing;
a testing system that performs a medical test on the media sampled from patients, each test instance having an outcome, wherein
the testing system comprises a high performance liquid chromatography system including a column holding a stationary medium,
the high performance liquid chromatography system separating components of the sampled media as the sampled media flows through
the column, and the high performance liquid chromatography system including a sensor that senses indications that the separated
components are passing the sensor, and wherein the sampled media is blood and the test measures the level of HbA1c hemoglobin
in the blood;

a reader that obtains, for each of a plurality of test instances, an identifier of the particular patient from which the media
was sampled by reading the identifier from a container that holds the media;

a processor that receives the output of the sensor and determines the outcome for each respective test and that causes test
outcomes to be stored in a mass storage memory in association with their respective patient identifiers; and

a computerized retrieval system that enables a user to specify a particular patient identifier and cause the computerized
retrieval system to retrieve from the mass storage memory test outcomes associated with the particular patient identifier.

US Pat. No. 9,766,261

LOW COST OPTICAL HIGH SPEED DISCRETE MEASUREMENT SYSTEM

Bio-Rad Laboratories, Inc...

1. A method for determining a velocity, an inflation rate, or both of a droplet passing through a microfluidic channel formed
of a translucent material, the method comprising:
illuminating the droplet with a first flash of light emitted by a multicolor light source, wherein the multicolor light source
is configured to emit light in two or more different wavelength bands and as temporally separated flashes, wherein each flash
comprises light of only one of said wavelength bands, and wherein the first flash of light comprises light of a first wavelength
band;

acquiring a first image of the droplet when the droplet is illuminated with the first flash of light with a detector configured
to detect and distinguish between light in said wavelength bands, wherein the multicolor light source, the detector, and the
microfluidic channel are positioned such that light emitted by the multicolor light source is incident upon the microfluidic
channel, and the droplet passing through the microfluidic channel and illuminated by the multicolor light source is imaged
on the detector;

allowing an amount of time to pass;
illuminating the droplet with a second flash of light emitted by the multicolor light source, wherein the second flash of
light comprises light of a second wavelength band different from the first wavelength band;

acquiring a second image of the droplet when the droplet is illuminated with the second flash of light, wherein the first
image and the second image are acquired (i) using the detector and (ii) as part of the same video frame or the same photographic
exposure;

calculating a change in position of the droplet, a change in size of the droplet, or both, between the first image and the
second image by separating the first image from the second image in the video frame or the photographic exposure; and

dividing the change in position of the droplet, the change in size of the droplet, or both, by the amount of time, thereby
determining a velocity of the droplet, an inflation rate of the droplet, or both, respectively, passing through the microfluidic
channel between the first image and the second image.

US Pat. No. 9,658,220

HUMAN FACTOR XIII AS A NORMALIZATION CONTROL FOR IMMUNOASSAYS

Bio-Rad Laboratories, Inc...

1. A method for normalizing results, the method comprising:
(a) contacting a biological sample with one or more solid supports having binding members immobilized thereon that specifically
bind an analyte in the biological sample, under conditions promoting binding of the analyte to the binding members, the solid
supports being divided into subpopulations that are differentiable from each other by a differentiation parameter comprising
a characteristic that is independent of the binding members immobilized on the solid supports, the binding members immobilized
on each subpopulation binding to one analyte in the sample;

(b) contacting the biological sample with a solid support having human blood coagulation Factor XIII (hFXIII) immobilized
thereon, the solid support being differentiable from the solid supports having binding members immobilized thereon;

(c) contacting the solid supports with a binding agent having binding affinity for hFXIII and binding agents having binding
affinity for each of the analytes, under conditions promoting binding of the binding agents to the analytes on the solid supports;

(d) quantifying the binding agents bound to the solid supports, and correlating the quantity of binding agents with the differentiation
parameters to obtain values individually representative of levels of the analytes and of hFXIII; and

(e) normalizing the values of the levels representative of the analytes to the value representative of hFXIII, thereby correcting
for variations and decreasing the coefficient of variation in sample processing or biological matrix effects during performance
of the method, wherein the normalizing comprises dividing the values of the levels representative of the analytes by the value
representative of hFXIII.

US Pat. No. 9,458,511

MULTIPLEXED DIGITAL ASSAY FOR VARIANT AND NORMAL FORMS OF A GENE OF INTEREST

Bio-Rad Laboratories, Inc...

1. A method of performing a multiplexed digital assay, the method comprising:
forming partitions each including a portion of a same sample comprising nucleic acid, the sample including a first target
that is a reference sequence, a second target that is a variant sequence from a gene of interest, and a third target that
is a normal sequence from the gene of interest, wherein each partition of only a subset of the partitions contains the first
target and the second target, and wherein each partition of only a subset of the partitions contains the third target;

amplifying the first target, the second target, and the third target in the partitions;
collecting data from a plurality of the partitions for amplification of each of the targets, wherein partitions positive for
both the first target and the second target are not distinguishable from partitions positive for only the third target;

determining a level of the second target; and
determining a copy number of the third target with a level of the first target and a level of the third target.
US Pat. No. 9,213,034

MULTIPLEX IMMUNOASSAYS FOR HEMOGLOBIN, HEMOGLOBIN VARIANTS, AND GLYCATED FORMS

Bio-Rad Laboratories, Inc...

1. A competitive immunoassay method for quantifying a plurality of hemoglobin analytes comprising HbA1c and a hemoglobin variant and glycated form thereof, if present, in a single sample of blood cell lysate, said method comprising:
(a) incubating said sample with a population of beads in a common mixture, said population comprising a HbA1c subpopulation to detect HbA1c and one or more further subpopulations, each of which detects a hemoglobin variant, wherein:

(i) each bead of said HbA1c subpopulation has bonded thereto a fluorescent dye and each bead of said one or more subpopulations to detect a hemoglobin
variant has bonded thereto a fluorescent dye, which may be the same as the fluorescent dye bonded to said subpopulation to
detect HbA1c or may be a different dye that distinguishes each of the one or more subpopulations; and

(ii) each bead of said HbA1c subpopulation to detect HbA1c further has bonded thereto an anti-HbA1c antibody having selective binding affinity towards HbA1c and each bead of said one or more further subpopulations to detect a hemoglobin variant has bonded thereto a monoclonal antibody
that has selective binding affinity towards the variant and the glycated form of the variant to cause each analyte to bind
to a different bead subpopulation through the antibody bonded to said subpopulations, wherein:

one subpopulation of the one or more further subpopulations to detect a hemoglobin variant is an HbC subpopulation that detects
HbC, if present in the sample, and has bonded thereto an anti-HbC monoclonal antibody having selective binding affinity towards
HbC and glycated HbC; or

one subpopulation of the one or more further subpopulations to detect a hemoglobin variant is an HbS subpopulation that detects
HbS, if present in the sample, and has bonded thereto an anti-HbS monoclonal antibody having selective binding affinity towards
HbS and glycated HbS; and

(b) individually detecting the subpopulations of beads that have been incubated with said sample in accordance with step (a)
to quantify the HbA1c and hemoglobin variant in the sample using a competitive immunoassay, wherein the competitive immunoassay comprises further
incubating the population of beads with a plurality of hemoglobin protein antigens each hemoglobin protein antigen attached
to a solid support at a separate site, wherein the plurality of hemoglobin protein antigens comprises a HbA1c antigen, which
HbA1c antigen is an HbA1c peptide that selectively binds the anti-HbA1c antibody; and an HbC antigen, which HbC antigen is an HbC peptide that selectively binds the anti-HbC monoclonal antibody
or an HbS antigen, which HbS antigen is an HbS peptide that selectively binds the anti-HbS monoclonal antibody; and

(c) quantifying HbA1c and the hemoglobin variant in the sample by determining competition of HbA1c present in the sample with the HbA1c peptide attached to the solid support for binding to the HbA1c subpopulation of beads; and determining competition of HbC present in the sample with the HbC peptide attached to the solid
support for binding to the HbC subpopulation of beads or determining competition of HbS present in the sample with the HbS
peptide attached to the solid support for binding to the HbS subpopulation of beads, thereby quantifying the hemoglobin analytes.

US Pat. No. 9,784,746

METHOD FOR STUDYING TRANSPORT OF AN AGENT ACROSS A BILAYER MEMBRANE IN BIOANALYTICAL SENSOR APPLICATIONS

Bio-Rad Laboratories, Inc...

1. A method comprising:
contacting a bilayer structure with at least one agent to be analyzed, the bilayer structure being tethered to at least one
metal surface and enclosing a detection volume;

detecting a change in refractive index only in the detection volume enclosed by the bilayer structure resulting from transportation
of the at least one agent across a membrane of the bilayer structure; and

determining a rate at which the at least one agent is transported across the bilayer structure using the detected change in
the refractive index of the detection volume.

US Pat. No. 9,738,683

ENHANCED FUNCTIONALITY AND DELIVERY OF A PROTEIN FROM A POROUS SUBSTRATE

Bio-Rad Laboratories, Inc...

1. A method of storing a protein, the method comprising:
providing a porous substrate and a protein aggregation modifying agent;
contacting the porous substrate in the presence of the protein aggregation modifying agent with a solution containing a protein;
and

reversibly immobilizing the protein on the porous substrate, wherein the reversible immobilization comprises drying the porous
substrate after contacting the porous substrate in the presence of the protein aggregation modifying agent with the solution
containing the protein, wherein the protein aggregation modifying agent is a cyclodextrin,

thereby storing the reversibly immobilized protein on the porous substrate.

US Pat. No. 9,623,384

SYSTEM FOR TRANSPORTING EMULSIONS FROM AN ARRAY TO A DETECTOR

Bio-Rad Laboratories, Inc...

1. A system for performing a droplet-based assay, comprising:
a droplet transporter including an inlet end, a channel junction, a source of carrier fluid including oil, and a detection
region formed by a microfluidic channel, wherein the channel junction is connected to the inlet end, the source, and the detection
region, wherein the inlet end is configured to be placed into each well of an array of wells, with each well holding an emulsion
including a plurality of droplets disposed in a continuous phase, and wherein the droplet transporter is configured to (a)
drive travel of droplets and the continuous phase of each emulsion into the inlet end and from the inlet end to the channel
junction, (b) add carrier fluid from the source to the continuous phase at the channel junction, to spatially separate droplets
from one another, and (c) drive travel of the spatially separated droplets through the detection region from the channel junction;

a detector configured to collect data related to one or more analytes from individual droplets of each emulsion as such individual
droplets travel through the detection region; and

a processor programmed to analyze the data to determine (i) a number of droplets containing at least one copy of an analyte
and/or a number of droplets containing no copies of the analyte, and (ii) a concentration of the analyte using at least one
of the numbers.

US Pat. No. 9,523,116

MULTIPLEXED DIGITAL ASSAY WITH DATA EXCLUSION FOR CALCULATION OF TARGET LEVELS

Bio-Rad Laboratories, Inc...

1. A method of performing a multiplexed digital assay, the method comprising:
providing a mixture including a plurality of targets and reagents sufficient for amplification of each of the targets, wherein
the plurality of targets includes a first target;

forming partitions each including a portion of the mixture;
amplifying the plurality of targets in the partitions;
collecting data from a plurality of the partitions for amplification of each of the plurality of targets;
identifying partition populations in the data, each partition population having a different target content of the plurality
of targets; and

calculating with a processor a level of the first target from only a portion of the data, wherein the portion of the data
is collected from only a subset of the plurality of the partitions, wherein the subset of partitions excludes two or more
of the partition populations, and wherein the two or more excluded partition populations include at least a pair of excluded
partition populations that are not resolved from each other in the data.

US Pat. No. 9,792,548

SYSTEMS AND METHODS FOR BIOCHEMICAL DATA ANALYSIS

Bio-Rad Laboratories, Inc...

1. A method of biochemical data analysis, the method comprising:
receiving, at a computer system, a dataset for a plurality of biological samples, the dataset having a plurality of fields
for each biological sample, at least a portion of the dataset being obtained from experiments involving the biological samples,
wherein the dataset includes:

a plurality of first fields, each first field including a plurality of values, each value corresponding to a respective characteristic
of a respective biological sample, and

a plurality of second fields, each second field corresponding to a respective analyte and including a plurality of concentrations
of the respective analyte in the experiments, each concentration corresponding to a respective biological sample;

in response to a user selecting the dataset via a graphical user interface (GUI) page of the computer system, automatically
populating a drop-down box of the GUI page with the plurality of first fields, wherein the user uses an input device to select
the dataset in the GUI page;

displaying, on a display of the computer system, a list of the plurality of first fields in a first region of the GUI page
in response to the user selecting the drop-down box in the GUI page, wherein the user uses the input device to select the
drop-down box;

receiving, at the GUI page, a selection of a compare field from the list of the plurality of first fields, wherein the user
uses the input device to select the compare field;

in response to the user selecting the compare field, automatically identifying a plurality of values in the database for the
selected compare field;

identifying, by the computer system, subgroups of the biological samples in the dataset for statistical analysis based on
the identified plurality of values for the selected compare field, wherein biological samples of a subgroup have a same value
for the compare field;

in further response to the user selecting the dataset, automatically displaying, on the display of the computer system, a
list of the plurality of second fields in a second region of the GUI page;

receiving, at the GUI page, a selection of an analyte from the list of the plurality of second fields displayed on the display
of the computer system for statistical analysis, wherein the user uses the input device to select the analyte;

in response to the user selecting the analyte, retrieving a plurality of concentrations of the selected analyte from the dataset;
in response to the user selecting the analyte and the compare field, automatically displaying, on the display of the computer
system, information of the retrieved plurality of concentrations separated by subgroups to convey statistical information
for the selected analyte for each subgroup of the compare field, wherein the information of the retrieved plurality of concentrations
separated by subgroups is displayed within a third region of the GUI page to facilitate visual comparison;

receiving, at the GUI page, a selection of a control group from a control group list, wherein the user uses the input device
to select the control group; and

in response to the user selecting the control group, automatically displaying, on the display of the computer system, a visual
indication as to which subgroup corresponds to the control group.

US Pat. No. 9,736,388

NON-DESTRUCTIVE READ OPERATIONS WITH DYNAMICALLY GROWING IMAGES

Bio-Rad Laboratories, Inc...

1. A method of capturing digital images of a specimen in a chemical reaction comprising:
capturing, by an imaging device, a series of short exposures of light emissions from the specimen over a period of time during
the reaction, wherein captured images of the specimen grow dynamically over the period of time as the number of exposures
increases,

wherein the series of short exposures is captured using an array of pixels of an image sensor of a digital imaging device
configured to perform continuous non-destructive read operations to read out a set of signals representing non-destructive
read images of the specimen from the array of pixels of the image sensor, and wherein reading out the final set of signals
is delayed until the end of the period of time to reduce read noise in the set of non-destructive read images;

monitoring, by the imaging device, the sets of signals representing the non-destructive read images read out from the image
sensor;

discontinuing capturing images of the specimen, by the imaging device, upon receiving a command, wherein the command is generated
based on the monitored signals; and

increasing a dynamic range of the captured images of the specimen by combining data from shorter-time read images of the set
of non-destructive read images for brighter areas of the specimen with data from longer-time read images of the set of non-destructive
read images for dimmer areas of the specimen.

US Pat. No. 10,161,947

USING PATIENT RISK IN ANALYSIS OF QUALITY CONTROL STRATEGY FOR LAB RESULTS

Bio-Rad Laboratories, Inc...

1. A method of analyzing a quality control (QC) strategy for an instrument used to measure an analyte in patient biological samples, the method comprising:obtaining, by a computer system, a non-failure probability of error PE(0) of the instrument for the analyte and an average number of patient results reported between failures (MPBF) of the instrument;
obtaining, by the computer system, information of the QC strategy, including:
an interval between QC events that monitor the instrument using a reference sample; and
a number of reference samples tested at each quality control event;
calculating, by the computer system, an expected number of unreliable final results E(Nuf) for the QC strategy;
obtaining, by the computer system, a probability Ph|u that an unreliable final result leads to patient harm;
computing, by the computer system, a proportion of patient results that are incorrect due to a failure of the instrument, the proportion including E(Nuf) divided by MPBF;
computing, by the computer system, a combined probability of incorrect results by adding PE(0) and the proportion;
computing, by the computer system, a predicted probability of harm by multiplying the combined probability and the probability Ph|u;
outputting, by the computer system, an assessment of the predicted probability of harm compared to an acceptable probability of harm, wherein the acceptable probability of harm is dependent on a severity of harm resulting from an incorrect result;
in response to the assessment indicating that the predicted probability of harm is lower or greater than the acceptable probability of harm, modifying the QC strategy by modifying the interval between QC events, the number of reference samples tested at each QC event, or both; and
conducting the QC events on the instrument according to the modified QC strategy.
US Pat. No. 9,885,728

VALUE ASSIGNMENT FOR CUSTOMIZABLE QUALITY CONTROLS

Bio-Rad Laboratories, Inc...

1. A method of preparing a quality control for an analyte, the method comprising:
applying a function to a desired reported concentration of an analyte in a quality control to return a target concentration
of the analyte in the quality control, wherein the desired reported concentration is a concentration value desired to be obtained
from a measurement procedure, wherein the measurement procedure is a technique or system for detecting a concentration of
an analyte, and wherein the function is derived by applying regression analysis to analyte concentration data previously obtained
using the measurement procedure;

providing one or more solid beads containing the analyte, and a base matrix, wherein the solid beads are water-soluble spheres
or pellets, and wherein the base matrix is a material substantially devoid of the analyte;

determining a number of solid beads and a volume of the base matrix needed to prepare the quality control with the target
concentration of the analyte; and

dissolving the number of beads in the volume of the base matrix, thereby preparing the quality control for the analyte.
US Pat. No. 9,797,902

HUMAN FACTOR XIII AS A NORMALIZATION CONTROL FOR IMMUNOASSAYS

Bio-Rad Laboratories, Inc...

1. A kit for determining whether a subject is infected with HIV or a stage of infection of a subject so infected, said kit
comprising a normalization factor immobilized on a solid support, and a plurality of binding members, each binding member
immobilized on a solid support, said plurality comprising:
(i) anti-p24 antibody,
(ii) HIV-1 envelope protein gp-160,
(ii) HIV-2 SPOH (RVTAIEKYLQDQARLNSWGCAFRQVC (SEQ ID NO:5)), and
(iv) HIV-O AFR (LNQQRLLNSWGCKGRLVCYTSV (SEQ ID NO:2));
said solid supports further bearing differentiation parameters selected such that all said supports bearing any one binding
member of said plurality are differentiable from all said supports bearing other binding members of said plurality.