Parkhomchuk Dmitri, Borodina Tatiana, Amstislavskiy Vyacheslav, Banaru Maria, Hallen Linda, Krobitsch Sylvia, Lehrach Hans, Soldatov Alexey
Max Planck Institute for Molecular Genetics, Department of Vertebrate Genomics, Ihnestr. 73, 14195 Berlin, Germany.
Nucleic Acids Res. 2009 Oct;37(18):e123. doi: 10.1093/nar/gkp596. Epub 2009 Jul 20.
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.
高通量互补DNA测序（RNA测序）是用于全转录组分析的强大工具，可提供有关转录本表达水平和结构的信息。然而，确定转录本的极性并因此识别哪条链被转录是困难的。在此，我们提出了一种简单的cDNA测序方案，该方案保留了有关转录本方向的信息。使用酿酒酵母和小鼠脑转录组作为模型，我们证明了解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，并能够研究启动子相关转录和反义转录。我们获得的转录图谱可在线获取。
