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Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

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Published on: April 17, 2015

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

Fumiki Katsuoka¹, Junji Yokozawa¹, Kaoru Tsuda¹

¹Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai 980-8573, Japan.

Analytical Biochemistry

|September 1, 2014

Summary

Quantitative MiSeq (qMiSeq) efficiently quantitates numerous libraries for Illumina HiSeq sequencers. This method uses index ratios from pooled libraries sequenced on a MiSeq to determine optimal HiSeq run conditions.

Keywords:

Library QC Library quantitation Next-generation sequencing

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Area of Science:

Genomics
Next-Generation Sequencing

Background:

Accurate library quantitation is essential for maximizing data output on Illumina HiSeq sequencers.
Existing methods can be time-consuming or less efficient for large numbers of samples.

Purpose of the Study:

To introduce a novel, efficient method for quantifying multiple sequencing libraries simultaneously.
To adapt the Illumina MiSeq platform for high-throughput library quantitation.

Main Methods:

Developed quantitative MiSeq (qMiSeq), a method utilizing the Illumina MiSeq platform.
Denatured and pooled 96 dual-index libraries with control samples.
Sequenced pooled libraries on MiSeq and analyzed observed index ratios.

Main Results:

Relative library concentrations were accurately determined using index ratios.
qMiSeq data enabled precise adjustments for Illumina HiSeq run conditions.
The method proved efficient for quantitating a large batch of libraries.

Conclusions:

qMiSeq offers an efficient and scalable solution for library quantitation prior to HiSeq sequencing.
This method simplifies and optimizes the pre-sequencing workflow.
qMiSeq facilitates high data output by ensuring proper library loading.