A kind of micro-degradation genomic DNA methylation library sequencing method and its kit

A genome and methylation technology, applied in the field of genomics and molecular biology, can solve problems such as undetectable, achieve the effect of low initial amount, reduce preference, and improve reaction efficiency
CN107858409BActive Publication Date: 2021-05-04SHENZHEN E GENE TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
SHENZHEN E GENE TECH
Publication Date
2021-05-04

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Abstract

The invention discloses a method for building and sequencing a micro-degraded genomic DNA methylation library. The invention tests different library building technologies and optimizes the reaction process to establish a methylation library that can detect as little as 1ng of severely degraded genomic DNA. technology. By testing different enzyme combinations, the end repair of degraded DNA, the addition of A at the 3-end, and the repair of DNA double-strand gaps due to degradation were completed in the same reaction system; by optimizing the reaction system, the sequencing adapters were connected in the original reaction solution; After further testing the biotin labeling of the adapter, through the specific binding of biotin and streptomycin magnetic beads, the capture of the genome fragments of the ligated adapter before the subsequent sulfite treatment was realized, and the loss of the target fragment was reduced; in addition, the establishment of sulfite Salt reaction system and reaction time reduce DNA damage during processing, ensure methylation conversion rate, and realize methylation library construction and sequencing of micro-degraded samples.
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Description

technical field

[0001] The invention relates to the technical fields of genomics and molecular biology, in particular to a method for building a library and sequencing a micro-degraded genome DNA methylation. Background technique

[0002] DNA methylation 5mC is a process in which the methyl group in the methyl donor S-adenoylmethionine (SAM) molecule is transferred to DNA cytosine mediated by methyltransferase (DNAmethyltransferase, MTase). In eukaryotes, DNA methylation mainly occurs at the 5-carbon position of the CpG dinucleotide cytosine, forming 5-methylcytosine (5mC). The methylation modification of DNA is involved in animal embryonic development, gene imprinting and X chromosome inactivation, and plays an important role in the regulation of gene expression. The disorder of DNA methylation modification provides new ideas for the exploration of abnormal embryonic development and the pathogenesis of cancer. In order to meet different scientific research needs, in the p...

Claims

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