Scalable transcriptome preparation for massive parallel sequencing.
basic_science · Level V
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- Record sourced from PubMed, PMID 21760920.
- Also identified by DOI 10.1371/journal.pone.0021910 and PMC identifier PMC2881125.
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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.