Construction method for next-generation sequencing library based on single-stranded connector and application thereof

A sequencing library and construction method technology, applied in the field of biomedicine, can solve problems such as loss of open area information, inability to detect DNA fragment sequences, etc., and achieve the effects of facilitating comparison and analysis, eliminating deviations, and simplifying library construction operations
CN107586835AActive Publication Date: 2018-01-16SOUTHEAST UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
SOUTHEAST UNIV
Publication Date
2018-01-16

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Abstract

The invention discloses a construction method for a next-generation sequencing library based on a single-stranded connector and application thereof. The method comprises the following steps: (1) carrying out denaturation on a double-stranded DNA (Deoxyribonucleic Acid) or RNA (Ribonucleic Acid) / DNA heterozygote segment to generate single-stranded DNA; (2) connecting one single-stranded connector with a 3' end of the single-stranded DNA; (3) extending the single-stranded DNA connected with the single-stranded connector by utilizing DNA polymerase to generate double-stranded DNA; (4) connectingthe other end of the double-stranded DNA with a T connector or a Tn5 label connector; (5) carrying out PCR (Polymerase Chain Reaction) amplification on the double-stranded DNA with two ends connectedwith the connectors to form a DNA library capable of sequencing of a next-generation sequencing technology. The method disclosed by the invention can be used for constructing the next-generation sequencing library and determining a DNA sequence and also can be used for identifying a chromatin open region, carrying out gene expression detection, carrying out trace nucleic acid amplification and thelike; the method is a novel method which belongs to the field of nucleic acid detection and analysis and has various functions and wide application value.
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Description

technical field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for constructing a next-generation sequencing (NGS) library based on Singlestrand Adapter Library Preparation (SALP) and its application. Background technique

[0002] Since the emergence of next-generation sequencing (NGS) on the market in 2005, this technology has changed the way we think about scientific research methods in the fields of basic, applied and clinical research. With the continuous development of new methods and computing power, NGS platforms have driven the explosion of biological knowledge in the past few years. As the most important application of NGS, the resequencing of the human genome has greatly deepened our understanding of the relationship between genetic diversity, health and disease. The biggest difference between NGS and traditional Sanger sequencing is that NGS needs to prepare a sequencing library. With the increasing data...

Claims

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