An efficient quantitation method of next-generation sequencing libraries by using MiSeq sequencer

Fumiki Katsuoka, Junji Yokozawa, Kaoru Tsuda, Shin Ito, Xiaoqing Pan, Masao Nagasaki, Jun Yasuda, Masayuki Yamamoto

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

Library quantitation is a critical step to obtain high data output in Illumina HiSeq sequencers. Here, we introduce a library quantitation method that uses the Illumina MiSeq sequencer designated as quantitative MiSeq (qMiSeq). In this procedure, 96 dual-index libraries, including control samples, are denatured, pooled in equal volume, and sequenced by MiSeq. We found that relative concentration of each library can be determined based on the observed index ratio and can be used to determine HiSeq run condition for each library. Thus, qMiSeq provides an efficient way to quantitate a large number of libraries at a time.

Original languageEnglish
Pages (from-to)27-29
Number of pages3
JournalAnalytical Biochemistry
Volume466
DOIs
Publication statusPublished - 2014 Dec 1

Keywords

  • Library QC
  • Library quantitation
  • Next-generation sequencing

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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