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深度探索RNA测序的结果

Iso-Seq方法能够对整个转录组或靶向基因的cDNA测序进行深度分析。所产生的全长mRNA转录本序列可通过一个简单的分析流程轻松获得。Iso-Seq分析还具有包括发现新的转录本和基因、鉴定融合基因以及注释异构体和可变剪接事件的能力。

适用于RNA测序分析的SMRT Analysis算法

Iso-Seq方法

利用单分子实时(SMRT)测序所产生的长序列,Iso-Seq方法可提供从5’端到3′ polyA尾巴、跨越整个转录本异构体的序列。准确的全长转录本序列的生成可显著简化分析——因为不需要使用短的RNA-Seq读取序列的容易出错的组装的转录本重建来推断异构体。

适用于RNA测序分析的SMRT Analysis应用程序

RS_IsoSeq.1

该应用程序适用于分析SMRT测序技术生成的数据,能够对转录本和剪接变体进行功能鉴定。

Iso-Seq分析运行可选择从头开始(de novo)或基于参考序列的模式运行。
它包括三个主要步骤:

  1. 分类:从PacBio系统(或SMRT Cell)运行中提取插入片段的序列;去除cDNA引物和poly-A;然后将插入片段的读取序列分成嵌合或非嵌合、全长或非全长的序列。
  2. 聚类:利用迭代聚类和错误纠正(ICE)算法,根据分类的读取序列预测新发的转录本一致性异构体。
  3. 映射:利用GMAP,将分类的读取序列和预测的一致性异构体与用户指定的参考序列进行比对。

更多资源

  1. SMRT Analysis Iso-Seq概述视频
  1. SMRT Analysis Iso-Seq生物信息学教程

     

    若有意了解如何运用这种创新的软件来支持您的研究工作,请联系我们