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Old 07-09-2011, 02:10 AM   #1
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Default RNA-Seq: Whole-transcriptome RNAseq analysis from minute amount of total RNA.

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Whole-transcriptome RNAseq analysis from minute amount of total RNA.

Nucleic Acids Res. 2011 Jul 6;

Authors: Tariq MA, Kim HJ, Jejelowo O, Pourmand N


RNA sequencing approaches to transcriptome analysis require a large amount of input total RNA to yield sufficient mRNA using either poly-A selection or depletion of rRNA. This feature makes it difficult to miniaturize transcriptome analysis for greater efficiency. To address this challenge, we devised and validated a simple procedure for the preparation of whole-transcriptome cDNA libraries from a minute amount (500 pg) of total RNA. We compared a single-sample library prepared by this Ovation® RNA-Seq system with two available methods of mRNA enrichment (TruSeq™ poly-A enrichment and RiboMinus™ rRNA depletion). Using the Ovation® preparation method for a set of eight mouse tissue samples, the RNA sequencing data obtained from two different next-generation sequencing platforms (SOLiD and Illumina Genome Analyzer IIx) yielded negligible rRNA reads (
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