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

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

Tohoku University

Research output: Contribution to journal › Article › peer-review

36 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 language	English
Pages (from-to)	27-29
Number of pages	3
Journal	Analytical Biochemistry
Volume	466
DOIs	https://doi.org/10.1016/j.ab.2014.08.015
Publication status	Published - 2014 Dec 1

Keywords

Library QC
Library quantitation
Next-generation sequencing

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10.1016/j.ab.2014.08.015

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title = "An efficient quantitation method of next-generation sequencing libraries by using MiSeq sequencer",

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",

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

note = "Funding Information: The authors thank the anonymous referees for their constructive suggestions and comments. This work was supported by the MEXT Tohoku Medical Megabank Project . We thank all members of the Tohoku Medical Megabank Organization. Publisher Copyright: {\textcopyright} 2014 Elsevier Inc.",

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doi = "10.1016/j.ab.2014.08.015",

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AU - Katsuoka, Fumiki

AU - Yokozawa, Junji

AU - Tsuda, Kaoru

AU - Ito, Shin

AU - Pan, Xiaoqing

AU - Nagasaki, Masao

AU - Yasuda, Jun

AU - Yamamoto, Masayuki

N1 - Funding Information: The authors thank the anonymous referees for their constructive suggestions and comments. This work was supported by the MEXT Tohoku Medical Megabank Project . We thank all members of the Tohoku Medical Megabank Organization. Publisher Copyright: © 2014 Elsevier Inc.

PY - 2014/12/1

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N2 - 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.

AB - 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.

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KW - Next-generation sequencing

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

Abstract

Keywords

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