Polynucleotide barcode generation
Assignee10x Genomics, Inc.
InventorsBenjamin Hindson, Mirna Jarosz, Paul Hardenbol, Michael Schnall-Levin, Kevin Ness, Serge Saxonov
Life of this patent
Estimating the term…
Abstract
The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing. In some cases, this disclosure provides methods for the generation of polynucleotide barcode libraries, and for the attachment of such polynucleotides to target polynucleotides.
Claims40
A method of synthesizing a library of polynucleotides comprising barcode sequences, said method comprising:
- synthesizing a plurality of polynucleotides comprising barcode sequences;
- separating said polynucleotides into a plurality of partitions, thereby generating partitioned polynucleotides, wherein each partition comprising a polynucleotide comprises multiple copies of the same polynucleotide comprising a barcode sequence prior to amplifying said partitioned polynucleotides;
- amplifying said partitioned polynucleotides, thereby generating a library of amplified polynucleotides, comprising at least about 100,000 different barcode sequences;
- wherein said amplified polynucleotides are attached to a bead;
- and
- isolating partitions comprising amplified polynucleotides.
A method of synthesizing a library of polynucleotides comprising barcode sequences, said method comprising:
- synthesizing a plurality of polynucleotides comprising barcode sequences;
- separating said polynucleotides into a plurality of partitions, thereby generating partitioned polynucleotides;
- amplifying said partitioned polynucleotides, using an RNA primer thereby generating amplified polynucleotides;
- optionally exposing said amplified polynucleotides to RNase H;
- and
- isolating partitions comprising amplified polynucleotides.
A method of synthesizing a library of polynucleotides comprising barcode sequences, said method comprising:
- synthesizing a plurality of MALBAC primer polynucleotides comprising barcode sequences;
- separating said polynucleotides into a plurality of partitions, thereby generating partitioned polynucleotides;
- amplifying said partitioned polynucleotides, thereby generating amplified polynucleotides;
- and
- isolating partitions comprising amplified polynucleotides.
Dependencies are inferred from the claim text.