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Chemical method for detecting 5-methylcystein in DNA

A technology of methylcytosine, chemical method, applied in the field of chemistry to detect 5-methylcytosine in DNA

Inactive Publication Date: 2013-02-06
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] However, the accuracy of these methods for detecting 5-methylcytosine needs to be further improved

Method used

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  • Chemical method for detecting 5-methylcystein in DNA
  • Chemical method for detecting 5-methylcystein in DNA
  • Chemical method for detecting 5-methylcystein in DNA

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Embodiment 1

[0033] The invention adopts the DNA sequence fragment of the promoter region in gastric cancer and cervical cancer to analyze, after reacting with p-nitrobenzenesulfonyl chloride sodium salt and m-nitrobenzenesulfonyl bromide sodium salt, and then treating with piperidine , the position of 5-methylcytosine in the DNA sequence can be clearly seen.

[0034] 1) Preparation of p-nitrobenzenesulfonyl chloride sodium salt

[0035] Add 0.8g of solid sodium hydroxide and 2mmol of p-nitrobenzenesulfonamide to 2ml of aqueous solution, stir for 5min, add 3ml of sodium hypochlorite solution, stir at room temperature for 12h, filter, wash the obtained solid with cold water for 5 times, and then reweight with water Crystallized to obtain pale yellow crystals. The yield was 83%. 1 H-NMR (300MHz, d 6 -DMSO)δ=8.27(d,J=7.5H z ,2H),7.88(d,J=7.5H z , 2H). 13 C-NMR (75MHz, d 6 -DMSO)δ=156.72,153.18,133.55,128.80.

[0036] 2) preparation of m-nitrobenzenesulfonyl bromide sodium salt

[003...

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Abstract

The invention relates to a chemical method for detecting 5-methylcystein in DNA (Deoxyribose Nucleic Acid). Two compounds adopted in the method are substituted chloramine-B hydrate and substituted bromoamine-B hydrate. The method comprises the detection steps of allowing the substituted chloramine-B hydrate to react with cytosine in the fluorescently-labeled DNA, then allowing the substituted bromoamine-B hydrate to react with the cytosine and the 5-methylcystein in the fluorescently-labeled DNA, treating with piperidine, and comparing two reacted DNA zones in DNA polyacrylamide denatured gel to clearly observe the positions and the number of the 5-methylcystein.

Description

technical field [0001] The invention relates to a chemical detection method of 5-methylcytosine in DNA. Background technique [0002] Epigenetics is defined as the study of inheritance of information on gene expression levels (quantitative changes). DNA methylation is the predominant form of epigenetics. [0003] DNA methylation has a great relationship with tumors. In 2003, three articles published in the same period in the American "Science" magazine have affirmed that the gene instability caused by the overall hypomethylation of DNA plays a causal role in the occurrence of tumors. In mouse models, it has now been demonstrated that chromosomal demethylation using mutations in DNA methyltransferase (DNMT) during embryonic to adult development can lead to genomic instability and lymphoma in mice. Generally speaking. The overall DNA methylation level of tumor cells is lower than that of normal cells, but some specific genes (such as tumor suppressor genes) CpG islands are...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 周翔王天露翁小成
Owner WUHAN UNIV
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