Transcriptome analysis by strand-specific sequencing of complementary DNA.

Transcriptome analysis by strand-specific sequencing of complementary DNA.
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DOI:
10.1093/nar/gkp596
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发表时间:
2009-10
影响因子:
14.9
通讯作者:
Soldatov A
Soldatov A
中科院分区:
生物学2区
文献类型:
--
作者:
Parkhomchuk D;Borodina T;Amstislavskiy V;Banaru M;Hallen L;Krobitsch S;Lehrach H;Soldatov A

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高通量互补DNA测序(RNA-Seq)是全转录组分析的有力工具,可提供有关转录本表达水平和结构的信息。然而,很难确定转录物的极性,因此很难确定哪条链被转录。在这里,我们提出了一个简单的cDNA测序协议,保留有关转录本的方向信息。使用酿酒酵母和小鼠脑转录组作为模型,我们证明,知道转录本的方向可以更准确地确定基因的结构和表达。它还有助于识别新基因,并使研究启动子相关和反义转录成为可能。我们获得的转录景观可在线获得。
High-throughput complementary DNA sequencing (RNA-Seq) is a powerful tool for whole-transcriptome analysis, supplying information about a transcript's expression level and structure. However, it is difficult to determine the polarity of transcripts, and therefore identify which strand is transcribed. Here, we present a simple cDNA sequencing protocol that preserves information about a transcript's direction. Using Saccharomyces cerevisiae and mouse brain transcriptomes as models, we demonstrate that knowing the transcript's orientation allows more accurate determination of the structure and expression of genes. It also helps to identify new genes and enables studying promoter-associated and antisense transcription. The transcriptional landscapes we obtained are available online.
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