Methods and compositions for analyzing nucleic acids
AssigneesThe Broad Institute, Inc., Massachusetts Institute of Technology, President and Fellows of Harvard College
InventorsJason Daniel Buenrostro, Sai Ma, Aviv Regev
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Abstract
Provided herein include methods and compositions for analyzing nucleic acid in individual cells. In some embodiments, the methods herein include generating, within individual cells, fragmented cellular genomic DNA and cDNA copies of cellular RNA molecules, barcoding the fragmented genomic DNA and the cDNA within each cell such that the genomic DNA and the cDNA from the same cell receive the same unique barcode sequence, isolating the barcoded genomic DNA and cDNA, and characterizing one or more features of the individual cells based, at least in part, on sequencing of the isolated barcoded genomic DNA and the cDNA.
Claims18
A method for single cell analysis of genomic DNA accessibility and RNA expression in a cell, comprising:
- a. fragmenting, within individual fixed cells, by a transposase, cellular genomic DNA;
- b. generating cDNA copies of cellular RNA molecules by reverse transcription;
- c. isolating the individual fixed cells in separate individual discrete volumes;
- d. barcoding with a primer pair and a volume-specific barcode, the primer pair and volume-specific barcode hybridizes to both fragmented cellular genomic DNA and cDNA such that the genomic DNA and the cDNA from the same cell receive the same unique cell barcode sequence;
- e. reverse crosslinking the individual fixed cells;
- f. isolating the barcoded genomic DNA and the cDNA;
- and
- g. characterizing one or more features of the individual cells based, at least in part, on sequencing of the isolated barcoded genomic DNA and the cDNA.
Dependencies are inferred from the claim text.