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

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, a...

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Bibliographic Details
Published in:Analytical biochemistry 2014-12, Vol.466, p.27-29
Main Authors: Katsuoka, Fumiki, Yokozawa, Junji, Tsuda, Kaoru, Ito, Shin, Pan, Xiaoqing, Nagasaki, Masao, Yasuda, Jun, Yamamoto, Masayuki
Format: Article
Language:English
Subjects:
Genomic Library
High-Throughput Nucleotide Sequencing - instrumentation
Library QC
Library quantitation
Next-generation sequencing
Sequence Analysis, DNA - methods
Sequence Analysis, DNA - standards
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Summary: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.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2014.08.015