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发明公开
CN105779439A 一种低起始量的高通量测序分析转录获得的RNA5’端信息的文库构建方法 无效 - 驳回

一种低起始量的高通量测序分析转录获得的RNA5’端信息的文库构建方法
摘要：
本发明属于基因组学技术领域，具体涉及一种能特异性获取5’ CAP序列信息的微量CAGE?seq文库构建方法，运用T4 PNK和TEX酶的组合作用，消除了总 RNA中5’端没有5’帽结构的RNA，保留了所有带有5’帽结构的RNA,设计相应的RT 引物和TS引物，在逆转录和模板置换过程中，富集RNA 5’端的信息，再通过PCR，引入测序引物和barcode序列。该方法避免繁琐的半抑制PCR过程，缩短建库时间，简化了实验流程；起始RNA用量较低，约100ng?1ug；是研究基因表达的有力方式，适用与基因调控网络研究。
摘要（英）：

The invention belongs to the technical field of genomics, and particularly relates to a micro CAGE-seq library construction method through which 5' CAP sequence information can be specifically acquired. Through the combined effect of T4 PNK and TEX, RNA without a 5' CAP structure at the 5' terminal in total RNA is eliminated, and all RNA with a 5' CAP structure is retained; a corresponding RT primer and a corresponding TS primer are designed, and RNA 5'-terminal information is enriched in the reverse transcription and template replacement processes; a sequencing primer and a barcode sequence are introduced through PCR. By means of the method, a tedious half-inhibiting PCR process is omitted, library construction time is shortened, and experiment procedures are simplified; the initial dose of RNA is low and is about 100 ng-1 microgram; the method is a powerful mode for studying gene expression and is suitable for studying gene regulatory networks.

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