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US 11,629,379 B2Granted
Single cell nucleic acid detection and analysis
AssigneePresident and Fellows of Harvard College
InventorsXiaoliang Sunney Xie, Katsuyuki Shiroguchi, Peter A. Sims, Tony Z. Jia
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Abstract
Methods and compositions for digital profiling of nucleic acid sequences present in a sample are provided.
Claims19
A composition comprising:
- a plurality of tagged cDNA molecules each labeled with (i) a copy number barcode sequence and (ii) a sample barcode sequence that identifies an isolated single cell providing RNA from which the cDNA was generated, wherein the plurality of tagged cDNA molecules are derived from a plurality of isolated single cells, wherein the sample barcode sequence identifies the isolated single cell providing RNA from which the tagged cDNA molecules were generated.
A composition of tagged cDNA molecules formed by a method comprising the steps:
- physically isolating a plurality of single cells to produce isolated single cells,
- releasing RNA from the isolated single cells, wherein the released RNA is physically separated from RNA released from other single cells,
- forming tagged cDNA from the released RNA, wherein the tagged cDNA comprises a (i) sample barcode sequence corresponding to the single cell and (ii) a copy number barcode sequence, and
- pooling the tagged cDNA derived from the different isolated single cells.
1 more element
Dependencies are inferred from the claim text.