公开(公告)号：US20100035249A1

公开(公告)日：2010-02-11

申请号：US12186009

申请日：2008-08-05

申请人： Yoshihide Hayashizaki ,  Piero Carninci ,  Mutsumi Katayama ,  Shintaro Katayama ,  Masayoshi Itoh ,  Matthias Harbers

发明人： Yoshihide Hayashizaki ,  Piero Carninci ,  Mutsumi Katayama ,  Shintaro Katayama ,  Masayoshi Itoh ,  Matthias Harbers

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6809 ,  C12Q1/6834 ,  C12Q1/6874 ,  C12Q2535/101 ,  C12Q2565/537 ,  C12Q2525/155

摘要： The present invention provides methods for the sequencing of all RNA species within an RNA sample, such as the RNA content obtained from a cell, a tissue, a living organism, or from an artificial source. RNA molecules within the samples are labeled in a RNA-specific manner prior to immobilization on a solid support. One label is used to mark the location of the RNA molecule on the solid support, whereas the second label is used to mark selectively the S′ end of full-length mRNA molecules. RNA molecules are sequenced while being bound to the solid support in one or more sequencing reactions, and sequences of individual RNA molecules can be forwarded to computational analysis for assembling sequence information from individual sequencing reads obtained from the same location on the solid support. Not only unsupervised expression profiling on a genome-wide scale, but also the direct analysis of RNA-RNA interactions become possible as revealed by the analysis of the sequencing information obtained along with genomic information.

摘要翻译： 本发明提供了用于对RNA样品中所有RNA物种进行测序的方法，例如从细胞，组织，活生物体或人造来源获得的RNA含量。 在固定在固体支持物上之前，将样品中的RNA分子以RNA特异性方式标记。 一个标签用于标记RNA分子在固体支持物上的位置，而第二个标记用于选择性标记全长mRNA分子的S'末端。 RNA分子在一个或多个测序反应中与固体支持物结合时进行测序，并且可将单个RNA分子的序列转发到计算分析，以从固体支持物上的相同位置获得的单独测序读数组装序列信息。 不仅在基因组范围内无监督的表达谱，而且通过分析与基因组信息一起获得的测序信息显示，RNA-RNA相互作用的直接分析也是可能的。