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.
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan. Electronic address: kfumiki@med.tohoku.ac.jp.
authors.forename: Fumiki
authors.initials: F
authors.lastname: Katsuoka
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.
authors.forename: Junji
authors.initials: J
authors.lastname: Yokozawa
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Tsuda
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Ito
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Pan
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Nagasaki
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Yasuda
authors.affiliation: 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: masiyamamoto@med.tohoku.ac.jp.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamamoto
mesh_terms: D015698:Genomic Library; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation*; D017422:Sequence Analysis, DNA / Q000379:methods* / Q000592:standards*
keywords: Library QC; Library quantitation; Next-generation sequencing