HIGH THROUGHPUT SEQUENCING OF MULTIPLE TRANSCRIPTS OF A SINGLE CELL

摘要

The present disclosure generally relates to sequencing two or more genes expressed in a single cell in a high-throughput manner. More particularly, the present disclosure relates to a method for high-throughput sequencing of pairs of transcripts co expressed in single cells (e.g., antibody VH and VL coding sequence) to determine pairs of polypeptide chains that comprise immune receptors.

主权项

1. A method comprising:
a) sequestering single cells and an mRNA capture agent into individual compartments; b) lysing the cells and collecting mRNA transcripts with the mRNA capture agent; c) isolating the mRNA from the compartments using the mRNA capture agent; d) performing reverse transcription followed by PCR amplification on the captured mRNA; and e) sequencing at least two distinct cDNA products amplified from a single cell.