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Method and kit for detecting DNA methylation

A kit and methylation technology, applied in the field of detection of DNA methylation, can solve the problems of cumbersome operation steps and achieve the effect of less DNA consumption, simple operation and good repeatability

Inactive Publication Date: 2016-01-06
BEIJING ACCB BIOTECH
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Problems solved by technology

However, the operation steps of this method are relatively cumbersome

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  • Method and kit for detecting DNA methylation
  • Method and kit for detecting DNA methylation
  • Method and kit for detecting DNA methylation

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[0047] Experimental procedure

[0048] Sample handling: Tissue genomic DNA was extracted using the QIAamp DNA Mini Kit (Qiagen). After the concentration was determined, the DNA was purified using a DNA purification kit (Tiangen Biochemical Technology Co., Ltd., Beijing) after treatment with MseI restriction endonuclease (at 37° C. for 2 hours). After measuring the DNA concentration, dilute the DNA to 10-50 ng / μL, denature at 95°C for 10 minutes, and place it on ice immediately. Take 1 μL and mix with 49 μL 1×blocking solution (PBS+3%BSA+0.05%TritonX-100), and place on ice.

[0049] Serum / plasma (200 μL) genomic DNA was extracted using the QIAamp DNA Blood Kit (Qiagen) and eluted with 45 μL of water. Denature at 95°C for 10 minutes and place on ice immediately. Add 5 μL of 10× blocking solution to mix and place on ice.

[0050] Sandwich method enzyme-linked immunosorbent assay (ELISA) experimental steps: The ELISA plate was coated with anti-methylcytosine antibody (Sant...

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Abstract

The invention relates to a method and kit for detecting DNA methylation. The method utilizes an enzyme-linked immunosorbent assay (ELISA) method to determine the whole methylation degree of DNA in cells, tissue and serum. The method has simple processes, good repeatability and a less DNA use amount (for detecting to ng level), is convenient for quantitative calculation and realizes high flux detection.

Description

technical field [0001] The invention relates to a method and a kit for detecting DNA methylation. Specifically, the present invention relates to a method and a kit for detecting DNA methylation by enzyme-linked immunosorbent assay (ELISA). Background technique [0002] DNA methylation occurs on the 5' carbon atom of cytosine in CpG dinucleotides, which is widely found in human cancers and is an important research object of epigenetics [1]. DNA methyltransferases (DNMTs) are responsible for the covalent modification of cytosine methyl groups [2]. In the DNMT family, DNMT1 is mainly responsible for maintaining the degree of DNA methylation, such as methylating the nascent DNA strand during DNA replication[3]; DNMT3a and DNMT3b are mainly responsible for methylating DNA from scratch Modification [4, 5]. DNA methylation information can be stably transmitted during cell division, and plays an important role in the development and function of organs, the inactivation of Z chrom...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 陈钊莫敏俐李晖李隽丁凤
Owner BEIJING ACCB BIOTECH
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