Quartz-Seq2: a high-throughput single-cell RNA-sequencing method that effectively uses limited sequence reads

Genome Biol. 2018 Mar 9;19(1):29. doi: 10.1186/s13059-018-1407-3.

Abstract

High-throughput single-cell RNA-seq methods assign limited unique molecular identifier (UMI) counts as gene expression values to single cells from shallow sequence reads and detect limited gene counts. We thus developed a high-throughput single-cell RNA-seq method, Quartz-Seq2, to overcome these issues. Our improvements in the reaction steps make it possible to effectively convert initial reads to UMI counts, at a rate of 30-50%, and detect more genes. To demonstrate the power of Quartz-Seq2, we analyzed approximately 10,000 transcriptomes from in vitro embryonic stem cells and an in vivo stromal vascular fraction with a limited number of reads.

Keywords: Cell sorter; Flow cytometry; High-throughput single-cell RNA-seq; Mesenchymal stem cell; Poly(A) tagging; Quartz-Seq; Stromal vascular fraction.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Embryonic Stem Cells / metabolism
  • Gene Expression Profiling / methods*
  • High-Throughput Nucleotide Sequencing / methods*
  • Mice
  • RNA-Directed DNA Polymerase
  • Sequence Analysis, RNA / methods*
  • Single-Cell Analysis / methods

Substances

  • RNA-Directed DNA Polymerase