RNA-Seq: a method for comprehensive transcriptome analysis.
RNA-Seq: a method for comprehensive transcriptome analysis.
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DOI：
10.1002/0471142727.mb0411s89
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发表时间：
2010-01-01
影响因子：
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通讯作者：
Snyder, Michael
中科院分区：
文献类型：
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作者：
Nagalakshmi, Ugrappa;Waern, Karl;Snyder, Michael
最近开发的一种称为RNA测序（RNA-Seq）的技术使用大规模并行测序，以比桑格测序和基于微阵列的方法更高的分辨率对基因组进行转录组分析。在RNA-Seq方法中，使用下一代测序技术直接对从感兴趣的RNA产生的互补DNA（cDNA）进行测序。然后可以将由此获得的读数与参考基因组进行比对，以构建全基因组转录组图谱。RNA-Seq已成功用于精确定量转录水平，确认或修改先前注释的基因的5 '和3'末端，以及绘制外显子/内含子边界。本单元描述了使用Illumina测序平台进行RNA-Seq的方案。
A recently developed technique called RNA Sequencing (RNA-Seq) uses massively parallel sequencing to allow transcriptome analyses of genomes at a far higher resolution than is available with Sanger sequencing- and microarray-based methods. In the RNA-Seq method, complementary DNAs (cDNAs) generated from the RNA of interest are directly sequenced using next-generation sequencing technologies. The reads obtained from this can then be aligned to a reference genome in order to construct a whole-genome transcriptome map. RNA-Seq has been used successfully to precisely quantify transcript levels, confirm or revise previously annotated 5' and 3' ends of genes, and map exon/intron boundaries. This unit describes protocols for performing RNA-Seq using the Illumina sequencing platform.