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Splice and sequencing library construction method

A sequencing library and molecular technology, applied in the fields of genetic engineering and molecular biology, can solve problems such as incorporation of adapters

Active Publication Date: 2013-02-27
WUHAN CONSIDERIN GENE & HEALTH TECH CO LTD
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  • Description
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Problems solved by technology

[0004] The purpose of the present invention is to provide a linker and a method for constructing a sequencing library, aiming at solving the problem of the prior art incorporating self-ligated products of the linker in the process of constructing a sequencing library

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  • Splice and sequencing library construction method

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Embodiment Construction

[0038] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0039] The present invention proposes a first typical embodiment, a linker, the linker is a complementary double-stranded nucleic acid molecule; only one strand of the double-stranded nucleic acid molecule contains at least one specific cleavage site for cleavage; the The specific cutting site is uracil nucleotide, P-S bond or deoxyinosine; the P-S bond is phosphorothioate bond.

[0040]The linker of this solution is suitable for joining with nucleic acid fragments of any length, especially for nucleic acid fragments that are not much different in length from the linker, for example: the size ratio of the nucleic acid segment to the linker is between 6:1 and 1:4, more preferably between Between 4:1 and 3:8, more preferably between 2:1 and 1:2. Bec...

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Abstract

The present invention relates to the field of genetic engineering and molecular biology, and provides a splice and a sequencing library construction method. The splice is complementary double-stranded nucleic acid molecule, wherein one strand of the double-stranded nucleic acid molecule contains at least one uracil nucleotide, a P-S bond or I, and the P-S bond is a phosphorothioate bond. The sequencing library construction method comprises: linking the splice and a fragmentation product, cutting the U, the P-S bond or the I on the splice so as to remove small nucleic acid fragments, and then using polymerase to carry out an extension reaction to obtain an extension product. With the splice and the sequencing library construction method, doping of the splice self-linking product in the constructed sequencing library can be avoided.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, and more specifically relates to a linker and a method for constructing a sequencing library. Background technique [0002] At this stage, DNA fragments used for high-throughput gene sequencing are first prepared into sequencing libraries. The preparation of the sequencing library includes the following steps. First, the sample genome is randomly cut to obtain a large number of DNA fragments, and then different adapters are connected to both ends of the DNA fragments. Finally, the sequencing library is obtained by amplifying the ligation products. In the process of building the library, the biggest problem is that the same adapter self-ligation and different adapters are easy to connect to each other when connecting the adapters, which leads to the mixed products of the adapter self-ligation in the constructed sequencing library, which interferes with the subsequent seque...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C40B50/06
Inventor 盛司潼
Owner WUHAN CONSIDERIN GENE & HEALTH TECH CO LTD
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