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获得一套更全面的转录本异构体

一个基因可能编码数量惊人的蛋白质,而每种蛋白质都具有独特的生物学功能。高等生物更是如此。短读长RNA测序(RNA-seq)的原理是将转录本异构体打断成较小的片段,然后利用生物信息学工具将其重新组装,这样就有可能错误组装,或无法完全捕获感兴趣基因的异构体多样性。

无需组装,捕获全长的转录本

PacBio的异构体测序(Iso-Seq)方法采用长读取序列来测序长达10 kb的转录本异构体。无论是广泛研究还是靶向分析,这种转录本多样性的分析都揭示了可变转录的频率和类型等关键信息,改善了基因组注释和基因发掘。

单分子实时(SMRT)测序增强了RNA测序,让您能够:

  • 开展直接的、基于证据的基因组注释
  • 发现新的基因和异构体,即使是研究透彻的样本
  • 鉴定各种基因模型中的启动子和剪接位点,以便了解基因调控
  • 在基因表达研究中,通过异构体水平的分辨率来改善RNA-seq定量的准确性
  • 区分重要的应激反应、发育和组织特异的异构体

工作流程:从RNA到全长转录本

Iso-Seq的样本制备

利用 PacBio系统 进行开展SMRT测序

  • 利用Sequel系统 简化样本制备流程和降低项目成本
  • 根据项目需求来扩大或缩小通量
    • 每个SMRT Cell 1M多达20Gb,或250到350 k的全长非嵌合读取序列 *
    • 分析单个SMRT Cell中的多个转录本
  • 探索Sequel系统上1到-2个SMRT Cell中的转录组
  • 提高测序深度,以便更全面地鉴定转录组
利用SMRT AnalysisPacBio DevNet开展数据分析

*读长,读取序列数,每个SMRT Cell产生数据,和其它测序表现结果会根据样品的类型/质量,插入片段长度以及其它因素产生变化

 

数据发布

 

蜂鸟和斑胸草雀大脑组织的Iso-Seq结果

利用Sequel系统生成的PacBio Iso-Seq数据集,探索转录组在禽类发声学习中的作用。

若想了解更多信息和访问数据,请查看我们的博客文章

Iso-Seq项目计算器

 

将您的动物/植物RNA测序项目告诉我们,即可获得一份在Sequel系统上开展Iso-Seq所需的时间和材料的估算。


研究聚焦:高粱基因组注释明显改善

Sorghum genome annotation

利用Iso-Seq方法开展的高粱转录组的剪接异构体分析大大改善了基因组注释,这种方法鉴定出77,000多个新的剪接异构体和2,100多个新基因。在这个例子中,科学家发现一个基因产生了13种新的可变剪接异构体,而之前的基因模型只包含一种异构体。深入了解这项研究:

Abdel-Ghany, S.E. et al. (2016) A survey of the sorghum transcriptome using single-molecule long reads. Nature Communications. 7, e11706.

若有意了解如何利用Iso-Seq分析转录组的复杂性,请联系我们

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