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发明公开
CN107794575A 用于Pacbio平台的DNA大片段文库构建方法和试剂盒 无效 - 驳回

用于Pacbio平台的DNA大片段文库构建方法和试剂盒
摘要：
本申请公开了一种用于Pacbio平台的DNA大片段文库构建方法和试剂盒。本申请用于Pacbio平台的DNA大片段文库构建方法，包括对打断的DNA大片段进行ExoVII酶处理；然后依序进行第一次DNA损伤修复、末端修复处理和加接头处理、外切酶ExoIII和ExoVII双酶切处理；然后，采用核酸电泳和片段回收系统对双酶切处理的产物进行片段分选；对片段分选产物进行第二次DNA损伤修复，即获得适用于Pacbio平台的DNA大片段文库。本申请的DNA大片段文库构建方法，整个建库过程无需使用Pacbio建库试剂盒，大大降低了建库成本，为第三代测序和Pacbio核酸测序平台的推广应用奠定了物质基础。
摘要（英）：

The application discloses a building method of DNA large-fragment library for Pacbio platform and an assay kit. The building method includes subjecting broken DNA large fragments to VII enzyme treatment, sequentially carrying out primary DNA damage repairing and terminal repairing treatment, linker splicing treatment, exonuclease ExoIII and ExoVII double-digest treatment, subjecting products obtained by double-digest treatment to fragment separation through nucleic acid electrophoresis and a fragment recovery system, subjecting products obtained by the fragment separation to secondary DNA damage repairing, so as to obtain the DNA large-fragment library adaptable to the Pacbio platform. According to the building method of the DNA large-fragment library, the whole building process is carriedout without Pacbio library-building assay kit, library building cost is lowered greatly, and a material foundation is laid for promotion and application of the third-generation sequencing and Pacbionucleic acid sequencing platform.

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