Genotyping of targeted loci with single-cell chromatin accessibility
AssigneesCornell University, New York Genome Center, Inc.
InventorsDan-Avi Landau, Robert Myers, Franco Izzo, Ronan Chaligne, Peter Smibert, Eleni Mimitou
Life of this patent
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Abstract
The present disclosure relates generally to methods, compositions, and systems for processing nucleic acids from individual cells or cell populations. The present invention enables direct sequencing of full-length RNA molecules with no amplification steps. The present invention allows determination of RNA modifications in highly specific cell populations of any tissue.
Claims32
A method of genotyping a locus, comprising:
- generating a plurality of biological particles, each biological particle comprising a polynucleotide;
- generating a plurality of partitions, each partition comprising one of the plurality of biological particles, a primer pair, and a barcoded bead;
- and
- wherein the primer pair comprises:
- a 5′ primer-adapter nucleic acid molecule comprising a nucleotide sequence that is complementary to a sequence located at the 3′ end of a locus within the polynucleotide, and
- a 3′ primer-adapter nucleic acid molecule comprising a nucleotide sequence that is complementary to a sequence located at the 5′ end of the locus,
- wherein the biological particle comprises genomic DNA and chromatin, and the genomic DNA is fragmented by a transposase to generate the plurality of genomic DNA fragments.
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35-36. (canceled)
The method of claim 24, further comprising amplifying the plurality of genomic DNA fragments, or derivatives thereof, by nucleic acid amplification generating a plurality of barcoded genomic DNA fragments.
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A partition comprising:
- a biological particle comprising a polynucleotide, a primer pair, and a barcoded bead;
- and
- wherein the primer pair comprises:
- a 5′ primer-adapter nucleic acid molecule comprising a nucleotide sequence that is complementary to a sequence located at the 3′ end of a locus within the polynucleotide, and
- a 3′ primer-adapter nucleic acid molecule comprising a nucleotide sequence that is complementary to a sequence located at the 5′ end of the locus.
66-103. (canceled)
A kit comprising partitioning fluid, a barcoded bead, and a primer pair comprising:
- a 5′ primer-adapter nucleic acid molecule comprising a nucleotide sequence that is complementary to a sequence located at the 3′ end of a specific locus within a polynucleotide, and
- a 3′ primer-adapter nucleic acid molecule comprising a nucleotide sequence that is complementary to a sequence located at the 5′ end of the specific locus.
105-116. (canceled)
Dependencies are inferred from the claim text.