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Composition for testing conversion efficiency of bisulfite-treated DNA in DNA methylation assays

A bisulfite and conversion efficiency technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of inconvenient inspection, uneconomical, expensive pyrosequencing equipment, etc.

Active Publication Date: 2022-07-29
BIOCHAIN BEIJING SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, pyrosequencing equipment is expensive, and most molecular biology laboratories do not have pyrosequencing equipment, and it is neither convenient nor economical to submit for inspection.

Method used

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  • Composition for testing conversion efficiency of bisulfite-treated DNA in DNA methylation assays
  • Composition for testing conversion efficiency of bisulfite-treated DNA in DNA methylation assays
  • Composition for testing conversion efficiency of bisulfite-treated DNA in DNA methylation assays

Examples

Experimental program
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Effect test

Embodiment 1

[0104] Example 1 Detection of conversion rate of bisulfite-treated DNA

[0105] 1) Use the whole blood DNA extraction kit (Boercheng (Beijing) Technology Co., Ltd.) to extract peripheral blood leukocyte genomic DNA (sample from Boercheng (Beijing) Technology Co., Ltd.), and artificially synthesize the sequence shown in SEQ ID NO: 2 of single-stranded DNA oligomers and prepared to a final concentration of 10 4 copies / µl of sample solution.

[0106] 2) Carry out bisulfite conversion treatment to above-mentioned DNA solution: use the bisulfite conversion kit within the validity period ((Boercheng (Beijing) Technology Co., Ltd., trade name is Septin9 gene methylation detection kit (PCR); Fluorescent probe method) Plasma treatment kit, production batch number 201610004), treated at 85°C for 0, 10, 20, 45, and 90 minutes respectively; at 60°C for 45 minutes; at the same time, use bisulfite conversion reagent beyond the expiration date Box ((Boercheng (Beijing) Technology Co., Lt...

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Abstract

The present invention provides a composition for detecting the conversion efficiency of bisulfite-treated DNA in DNA methylation detection, the composition comprising a reference nucleic acid, and primers and probes for detecting the reference nucleic acid . The present invention also provides a method for detecting the conversion efficiency of bisulfite-treated DNA in DNA methylation detection. The method of the present invention can effectively and accurately detect the transformation efficiency of bisulfite-treated DNA in DNA methylation detection, and can especially help to find out the transformation process such as insufficient transformation time, too low transformation temperature, and expired transformation reagents. possible problems.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a composition for detecting the conversion efficiency of bisulfite-treated DNA in DNA methylation detection, and also relates to a composition for detecting sub-sulfite in DNA methylation detection. A method and a kit for the transformation efficiency of DNA treated with bisulfate. Background technique [0002] DNA methylation is a regulatory mechanism of epigenetics. 5-methylcytosine is generated by methylating cytosine in CpG base combinations in specific regions of the genome sequence. The methylation modification of DNA cytosine is important in gene expression regulation, genomic imprinting, embryonic development, maintenance of normal It plays an extremely important role in epigenetic regulation of cell function and disease occurrence. [0003] Since 5-methylcytosine in methylated DNA has the same base pairing properties as cytosine in unmethylated DNA, it cannot be directly det...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6876C12N15/11
CPCC12Q1/6876C12Q2600/154
Inventor 马竣韩晓亮王建铭
Owner BIOCHAIN BEIJING SCI & TECH