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Detection method of folate metabolism related gene

A technology of folic acid metabolism and detection method is applied in the field of detection of folic acid metabolism-related genes, which can solve the problems of poor detection accuracy and slow speed.

Inactive Publication Date: 2017-10-17
安徽安龙基因科技有限公司
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a detection method for genes related to folic acid metabolism in order to overcome the defects of poor detection accuracy and slow speed in the prior art

Method used

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  • Detection method of folate metabolism related gene
  • Detection method of folate metabolism related gene
  • Detection method of folate metabolism related gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Pregnant woman Tang was tested for genes related to folic acid metabolism

[0063] (1) Using a buccal swab DNA extraction kit to extract DNA from Tang’s oral mucosal cells, ensure that the DNA concentration is 25ng / ul, 260 / 280=1.91;

[0064] (2), using SEQ NO 1 and SEQ NO 2 to amplify the MTHFR gene C677T; using SEQ NO 3 and SEQ NO 4 to amplify the MTHFR gene A1298C; using SEQ NO 5 and SEQ NO 6 to amplify the MTRR gene A66G. The DNA extracted from oral mucosal cells was used as a template for PCR reaction, and a 25uL system was taken, which included:

[0065] Template: 2uL, SEQ NO 1: 1uL, SEQ NO 2: 1uL, 10×MSP buffer: 2.5uL, 20mM dNTP: 2ul, Hot start taq: 0.3ul, deionized water: 16.2uL;

[0066] Template: 2uL, SEQ NO 3: 1uL, SEQ NO4: 1uL, 10×MSP buffer: 2.5uL, 20mM dNTP: 2ul, Hot start taq: 0.3ul, deionized water: 16.2uL;

[0067] Template: 2uL, SEQ NO 5: 1uL, SEQ NO 6: 1uL, 10×MSP buffer: 2.5uL, 20mM dNTP: 2ul, Hot start taq: 0.3ul, deionized water: 16.2uL;

[0068]...

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Abstract

The invention discloses a detection method of a folate metabolism related gene, which includes steps of extracting DNA from oral mucosa cell of a detector; amplifying MTHFR gene C677T by SEQ NO 1 and SEQ NO 2; amplifying MTHFR gene A1298C by SEQ NO 3 and SEQ NO 4; amplifying MTHFR gene A66G by SEQ NO 5 and SEQ NO 6; extracting DNA from oral mucosa cell as a template to perform PCR reaction; performing sepharose gel electrophoresis of 1% of ordinary PCR amplified product on three pairs of primers; recycling a strip of a kit recycling item from the 1% of agarose gel electrophoretic band by sepharose gel; performing sequence testing reaction; purifying the sequence testing reaction product and denaturing; testing sequence by an upper 3730 sequence tester; analyzing three SNP gene models of the sequence testing result by Chromas software. The method has the advantages of high detection accuracy, and convenience of use.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a detection method for genes related to folic acid metabolism. Background technique [0002] Folic acid, also known as pteroyl glutamic acid, is composed of three parts: pteridine nucleus, p-aminobenzoic acid and glutamic acid. Both humans and mammals can only absorb exogenous folic acid through the intestines and cannot synthesize folic acid by themselves. Folic acid is mainly absorbed by the upper part of the small intestine. The epithelial mucosal cells of the duodenum and jejunum contain folate reductase. Under the action of this enzyme, folic acid is methylated to generate dihydrofolate, and then methylated to generate tetrahydrofolate. Methyltetrahydrofolate is a carrier of one-carbon units and participates in the biosynthesis of important substances such as purine and pyrimidine that make up DNA and RNA. Folic acid metabolism also has high individual differences, and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6869C12Q1/6883C12Q2600/156C12Q2535/101
Inventor 韦玉军李航崔俊生苏军吴远航
Owner 安徽安龙基因科技有限公司
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