A highly sensitive and accurate gene expression analysis by sequencing ("bead-seq") for a single cell

Hiroko Matsunaga, Mari Goto, Koji Arikawa, Masataka Shirai, Hiroyuki Tsunoda, Huan Huang, Hideki Kambara*

*この研究の対応する著者

研究成果: Article査読

13 被引用数 (Scopus)

抄録

Analyses of gene expressions in single cells are important for understanding detailed biological phenomena. Here, a highly sensitive and accurate method by sequencing (called "bead-seq") to obtain a whole gene expression profile for a single cell is proposed. A key feature of the method is to use a complementary DNA (cDNA) library on magnetic beads, which enables adding washing steps to remove residual reagents in a sample preparation process. By adding the washing steps, the next steps can be carried out under the optimal conditions without losing cDNAs. Error sources were carefully evaluated to conclude that the first several steps were the key steps. It is demonstrated that bead-seq is superior to the conventional methods for single-cell gene expression analyses in terms of reproducibility, quantitative accuracy, and biases caused during sample preparation and sequencing processes.

本文言語English
ページ(範囲)9-16
ページ数8
ジャーナルAnalytical Biochemistry
471
DOI
出版ステータスPublished - 2015 2 15
外部発表はい

ASJC Scopus subject areas

  • 生物理学
  • 生化学
  • 分子生物学
  • 細胞生物学

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