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Scalable transcriptome preparation for massive parallel sequencing.

ABSTRACT:

Background

The tremendous output of massive parallel sequencing technologies requires automated robust and scalable sample preparation methods to fully exploit the new sequence capacity.

Methodology

In this study, a method for automated library preparation of RNA prior to massively parallel sequencing is presented. The automated protocol uses precipitation onto carboxylic acid paramagnetic beads for purification and size selection of both RNA and DNA. The automated sample preparation was compared to the standard manual sample preparation.

Conclusion/significance

The automated procedure was used to generate libraries for gene expression profiling on the Illumina HiSeq 2000 platform with the capacity of 12 samples per preparation with a significantly improved throughput compared to the standard manual preparation. The data analysis shows consistent gene expression profiles in terms of sensitivity and quantification of gene expression between the two library preparation methods.

SUBMITTER: Stranneheim H

PROVIDER: S-EPMC3131396 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Scalable transcriptome preparation for massive parallel sequencing.

Stranneheim Henrik H Werne Beata B Sherwood Ellen E Lundeberg Joakim J

PloS one 20110707 7

<h4>Background</h4>The tremendous output of massive parallel sequencing technologies requires automated robust and scalable sample preparation methods to fully exploit the new sequence capacity.<h4>Methodology</h4>In this study, a method for automated library preparation of RNA prior to massively parallel sequencing is presented. The automated protocol uses precipitation onto carboxylic acid paramagnetic beads for purification and size selection of both RNA and DNA. The automated sample preparat ...[more]

PMID: 21760920

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