Construction method, kit and application of plasma free DNA methylation detection library

A construction method and methylation technology, which is applied in the field of plasma cell-free DNA methylation detection library construction, can solve the problems that plasma cell-free DNA cannot construct a library, reduce the initial amount of library construction, improve library recovery efficiency, simplify The effect of the library building step
CN107541791AInactive Publication Date: 2018-01-05BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
Publication Date
2018-01-05
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a construction method, a kit and application of a plasma free DNA methylation detection library. The construction method comprises the following steps: S1 of extracting plasmafree DNA from a blood sample; S2 of performing tail end repair and 3' terminal A base addition on the plasma free DNA; S3 of connecting the tail ends of the plasma free DNA obtained in the S2 with C methylation modified connectors; S4 of performing bisulfite conversion on a product obtained in S3 and performing PCR expansion; S5 of performing magnetic bead purification on the PCR product of S4 andremoving small-fragment DNA which is not amplified in a non-specific mode and primer dimer; S6 of performing PCR amplification on a product of S5 and performing magnetic bead purification to obtain aplasma free DNA methylation detection library. By means of the technical scheme of the construction method disclosed by the invention, an initial amount of general microscale methylation library construction is reduced, library construction steps are simplified, library recycling efficiency is improved, and a foundation is established for sequentially screening tumor diagnosis markers through plasma free DNA methylation difference locus.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for constructing a plasma free DNA methylation detection library, a kit and an application thereof. Background technique

[0002] Plasma free circulating tumor DNA (circulating tumor DNA, ctDNA) refers to single-stranded or double-stranded DNA released into plasma by necrotic or apoptotic tumor cells, accounting for about 0.1%-1% of free DNA. The size of the ctDNA fragment is usually 160-180bp, with a half-life of 2 hours in the blood, and carries information such as SNP (point mutation), InDel (insertion deletion), CNV (copy number), fusion (fusion gene), methylation, etc. ctDNA flows continuously in the blood circulation system, which can reflect the current information of tumor patients in real time.

[0003] Compared with traditional detection methods, ctDNA detection has many advantages: 1) Convenient sampling: ctDNA detection only needs to draw blood to complete the a...

Claims

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