一种核酸的双接头单链环状文库的构建方法和试剂技术

一种核酸的双接头单链环状文库的构建方法和试剂，该方法包括：将核酸打断成核酸片段；连接第一接头序列；扩增得到两端具有第一接头序列的第一产物，其中引物序列上具有U碱基；用USER酶酶切第一产物，环化产生缺口；或引物序列上还具有切口酶识别序列，用USER酶酶切第一产物，环化并使用切口酶酶切产生切口；从切口或缺口处进行限制性切口/缺口平移反应；消化除去未发生限制性切口/缺口平移反应的部分；连接第二接头序列；扩增得到两端具有第二接头序列的第二产物；变性第二产物，并使用介导序列对单链核酸分子进行环化。该方法能够提高文库插入片段长度，并且无需切胶回收；单链核酸分子热变性后可以直接进行环化。

Construction method and reagent for double linked single strand ring Library of nucleic acid

Methods and reagents to construct a double joint circular single stranded nucleic acid library, the method comprises: nucleic acid broken into nucleic acid fragments; connecting the first joint sequence; amplification of the first product ends with a first joint sequence, the primer sequences with U bases; cut the first product with USER enzyme, cyclization to produce gap; or primer sequences have incision enzyme recognition sequence, cut the first product with USER enzyme, cyclization and use of enzyme generated incision incision; limited incision / nick translation reaction from incision or gaps; without restriction digestion to remove incision / nick translation reaction; connecting the second joint sequence amplification; second the product has second joints at both ends of the sequence; degeneration of second products, and the use of dielectric ring of single stranded nucleic acid molecules guide sequence. The method can increase the length of the inserted fragments of the library without the need for gel recovery, and the single stranded nucleic acid molecules can be directly cyclization after thermal denaturation.

【技术实现步骤摘要】
【国外来华专利技术】PCT国内申请，说明书已公开。

【技术保护点】
PCT国内申请，权利要求书已公开。

【技术特征摘要】
【国外来华专利技术】PCT国内申请，...

【专利技术属性】
技术研发人员：江媛，赵霞，阿莱克谢耶夫·安德烈，徳马纳克·拉多杰，章文蔚，蒋慧，
申请(专利权)人：深圳华大基因研究院，深圳华大基因科技有限公司，
类型：发明
国别省市：广东,44