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US 2020/0263232 A1Published application
Methods and systems to minimize barcode exchange
Assignee10x Genomics, Inc.
InventorsJason Bell, Josephine Lee, Corey Nemec, Francesca Meschi
Life of this patent
Estimating the term…
Abstract
Methods and compositions to minimize barcode exchange during the preparation of barcoded next-generation sequencing libraries prepared from a single cell. The methods utilize oligonucleotides containing a 3′-terminated blocking group or sequences that prevent amplification or extension.
Claims21
-83. (canceled)
A method for nucleic acid processing, comprising:
- (a) contacting a plurality of biological particles comprising template nucleic acid molecules with a plurality of transposase complexes comprising adapter sequences to generate, from said template nucleic acid molecules, a plurality of adapter-flanked fragments, wherein said plurality of biological particles is a plurality of cells or a plurality of cell nuclei;
- (b) partitioning said plurality of biological particles into a plurality of partitions, wherein a partition of said plurality of partitions comprises:
- (i) a biological particle of said plurality of biological particles, wherein said biological particle comprises an adapter-flanked fragment of said plurality of adapter-flanked fragments;
- (ii) a plurality of nucleic acid barcode molecules comprising a common barcode sequence;
- and
- (iii) a nucleic acid splint molecule comprising a first sequence complementary to said adapter sequence and a second sequence complementary to a sequence of said nucleic acid barcode molecule, wherein said nucleic acid splint molecule comprises a non-canonical base;
- and
- (c) using said adapter-flanked fragment, said nucleic acid barcode molecule, and said nucleic acid splint molecule to generate a barcoded nucleic acid molecule comprising a sequence of said adapter-flanked fragment and said common barcode sequence.
5 more elements
Dependencies are inferred from the claim text.