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

doi:10.1016/j.ab.2014.08.015

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

Anal Biochem. 2014 Dec 1:466:27-9. doi: 10.1016/j.ab.2014.08.015. Epub 2014 Aug 28.

Authors

Fumiki Katsuoka¹, Junji Yokozawa², Kaoru Tsuda², Shin Ito², Xiaoqing Pan², Masao Nagasaki², Jun Yasuda², Masayuki Yamamoto³

Affiliations

¹ Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan. Electronic address: [email protected].
² Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.
³ Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan; Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8575, Japan. Electronic address: [email protected].

PMID: 25173513
DOI: 10.1016/j.ab.2014.08.015

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.

Keywords: Library QC; Library quantitation; Next-generation sequencing.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Genomic Library*
High-Throughput Nucleotide Sequencing / instrumentation*
Sequence Analysis, DNA / methods*
Sequence Analysis, DNA / standards*