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    Droplet tagging contiguity preserved tagmented DNA
    12.
    发明授权
    Droplet tagging contiguity preserved tagmented DNA 有权

    公开(公告)号：US11685947B2

    公开(公告)日：2023-06-27

    申请号：US17236743

    申请日：2021-04-21

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Ronald Lebofsky ,  Jeremy Agresti

    IPC: C12P19/34 ,  C12Q1/6809 ,  G16B20/00 ,  C12Q1/6869

    CPC classification number: C12Q1/6809 ,  C12Q1/6869 ,  G16B20/00 ,  C12Q1/6869 ,  C12Q2521/301 ,  C12Q2563/149 ,  C12Q2563/159 ,  C12Q2565/514 ,  C12Q2565/537

    Abstract: Methods and compositions for maintaining DNA contiguity for sequencing is provided. For example, a plurality of partitions is provided comprising a bead, a forward primer oligonucleotide cleaved from the bead, the forward primer oligonucleotide having a bead-specific barcode and a 3′ end specific for and complementary to a first or second adaptor; a reverse primer oligonucleotide having a 3′ end complementary to the first or second adaptor, wherein the forward primer 3′ end and the reverse primer 3′ end are complementary to different adaptors selected from the first adaptor and the second adaptor; and fragments of genomic DNA reacted with an adapter-loaded tagmentase such that the DNA fragments comprise breakpoints in the fragments and an inserted adaptor at the break points, wherein the tagmentase binds the breakpoints to form linked DNA segments in the form of DNA segment-first adaptor tagmentase second adaptor-(DNA segment-first adaptor tagmentase second adaptor)n-DNA segment, where n is any integer and “-” indicates a covalent linkage.

    Multiple beads per droplet resolution
    16.
    发明授权
    Multiple beads per droplet resolution 有权

    公开(公告)号：US10730030B2

    公开(公告)日：2020-08-04

    申请号：US15400756

    申请日：2017-01-06

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Ronald Lebofsky ,  Nicholas Heredia

    IPC: B01J19/00 ,  C12Q1/6806

    Abstract: Methods of generating a nucleic acid signature for identifying particles associated in a partition are provided. In one aspect, the method comprises: partitioning a sample into a plurality of partitions comprising a particle comprising a solid support surface, the solid support surface having a plurality of oligonucleotide primers conjugated thereon, wherein the oligonucleotide primers comprise a barcode sequence, and wherein the partitions have 0, 1, or more than 1 particles per partition; providing in a partition a substrate comprising a barcode sequence or repeating clonal barcode sequences; and in the partition, associating a first particle conjugated to oligonucleotide primers comprising a first barcode sequence and a second particle conjugated to oligonucleotide primers comprising a second barcode sequence to a barcode sequence from the substrate, thereby generating a nucleic acid signature for the particles in the partition.

    SELECTIVE DIGITAL MULTIPLEXING
    18.
    发明申请
    SELECTIVE DIGITAL MULTIPLEXING 有权

    公开(公告)号：US20250043362A1

    公开(公告)日：2025-02-06

    申请号：US18793760

    申请日：2024-08-03

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： George Karlin-Neumann ,  Ronald Lebofsky ,  Mona Shahbazian ,  Mina Rostamza

    IPC: C12Q1/6886 ,  C12Q1/6806 ,  C12Q1/6809 ,  C12Q1/686 ,  G01N21/64

    Abstract: The invention provides methods for the detection of molecular targets by digital PCR (dPCR) with a blocker for non-target molecules. Each target is provided with a unique mixture of probes. The blocker binds to a non-target molecule and blocks it from contributing to fluorescence from partitions. Two or more colors of fluorescence intensity are read, and two colors of fluorescence intensity are plotted as a 2D plot. In the plot, different targets contribute well-resolved clusters. Each cluster in the plot essentially lies a long its own radius allowing for radial multiplexing. The use of a blocker selects against the detection of non-target molecules and improves the resolution of radial multiplexing, allowing multiple targets to be detected.

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