High-throughput sequencing library and its construction method

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...

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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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