Quantitative single-cell RNA-seq with unique molecular identifiers

Nat Methods. 2014 Feb;11(2):163-6. doi: 10.1038/nmeth.2772. Epub 2013 Dec 22.

Abstract

Single-cell RNA sequencing (RNA-seq) is a powerful tool to reveal cellular heterogeneity, discover new cell types and characterize tumor microevolution. However, losses in cDNA synthesis and bias in cDNA amplification lead to severe quantitative errors. We show that molecular labels--random sequences that label individual molecules--can nearly eliminate amplification noise, and that microfluidic sample preparation and optimized reagents produce a fivefold improvement in mRNA capture efficiency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Gene Expression Profiling
  • Gene Library
  • High-Throughput Nucleotide Sequencing / methods*
  • Mice
  • RNA, Messenger / genetics*
  • Sequence Analysis, RNA / methods*
  • Systems Biology

Substances

  • RNA, Messenger

Associated data

  • GEO/GSE46980