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High-throughput sequencing library and its construction method

A construction method and technology for sequencing libraries, applied in libraries, chemical libraries, nucleotide libraries, etc., can solve the problems of low sequencing efficiency and small amount of effective data, and achieve the effects of reducing invalid data, improving purity, and increasing effective amount

Active Publication Date: 2017-06-06
BEIJING NOVOGENE TECH CO LTD
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Problems solved by technology

[0018] The main purpose of the present invention is to provide a high-throughput sequencing library and its construction method, so as to improve the shortcomings of the library constructed in the prior art, such as small amount of effective data and low sequencing efficiency.

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  • High-throughput sequencing library and its construction method
  • High-throughput sequencing library and its construction method

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[0036] It should be noted that, in the case of no conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and examples.

[0037] Exonuclease in the present invention refers to the nuclease that acts on double-stranded DNA, has 3 ' → 5 ' exonuclease activity but does not have DNA polymerase activity. What the exonucleases applicable to the present invention have in common is that they all use double-stranded DNA as a substrate, and have the activity of excising single nucleotides one by one from the 3'-OH end without the activity of DNA polymerase , Nucleases that meet the above conditions are all within the protection scope of the present invention, and, in addition to having the above-mentioned activities, the exonucleases of the present invention can also have other relatively weak nuclease activities, such as B...

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Abstract

The invention discloses a high-throughput sequencing library and a construction method thereof. The construction method comprises the following steps: S1. acquiring a DNA fragment from a DNA-protein cross-linked complex; S2. enzymatically digesting the DNA fragment by virtue of exonuclease; and S3. constructing a library for the enzymatically digested DNA fragment so as to obtain the high-throughput sequencing library, wherein the exonuclease refers to nuclease which acts on double-stranded DNA only and has the activity of 3'->5' exonuclease rather than the activity of DNA polymerase. By replacing a klenow fragment which has the activity of the 3'->5' exonuclease and the activity of the DNA polymerase in the prior art by virtue of the exonuclease which has the activity of 3'->5' exonuclease and does not have the activity of DNA polymerase, the library construction method disclosed by the invention can be used for removing the marker in a non-target fragment more thoroughly, so that the purity of the obtained DNA fragment is greatly improved, and subsequently the effective quantity of subsequent sequencing data is increased.

Description

technical field [0001] The invention relates to the field of high-throughput sequencing, in particular to a high-throughput sequencing library and a construction method thereof. Background technique [0002] Chromosome is the carrier of genetic material, and its activity and function are determined by its structure and spatial conformation. Chromosome composition has been in a dynamic balance for a long time, that is, it is different throughout the cell cycle and even in different types of cells. This different composition and activity plays an important role in the morphological construction of chromosomes and the expression regulation and replication of genes. In 2002, Job Dekker et al. first invented this method for studying the spatial conformation of chromosomes, that is, the Chromosome Conformation Capture method, or the 3C method, which can be used to study the interaction frequency of any two genetic loci on chromosomes . The principle of the 3C method is: firstly...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/10C40B50/06C40B40/06
CPCC12Q1/6869C40B50/06C12Q2535/122
Inventor 蒋智王大伟李明洲李宗文朱海浩王苗英刘运超
Owner BEIJING NOVOGENE TECH CO LTD
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