scRNA-seq for Microcephaly Research [II]: Preparation of Single-Cell cDNA Libraries

Methods Mol Biol. 2023:2583:99-104. doi: 10.1007/978-1-0716-2752-5_9.

Abstract

Droplet-based single-cell RNA-seq (scRNA-seq) requires the addition of bar codes that mark the cell and transcript of origin. Cell-specific bar codes, typically added by cDNA elongation on beads, label each transcript derived from an individual cell. Transcript-specific bar codes serve as unique molecular identifiers and allow for the maintenance of transcript proportions after PCR-based amplification of cDNA. This chapter provides methods for generating bar-coded scRNA-seq libraries after droplet encapsulation.

Keywords: Drop-seq; Transcriptomic analysis; scRNA-seq.

MeSH terms

  • DNA, Complementary / genetics
  • Gene Library
  • Humans
  • Microcephaly*
  • Sequence Analysis, RNA / methods
  • Single-Cell Analysis* / methods

Substances

  • DNA, Complementary