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Kit and method for genotyping detection

A genotyping and kit technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high genotyping cost and cumbersome operation, and achieve short time-consuming, good repeatability, and optimized PCR Effect of Amplification Conditions

Inactive Publication Date: 2018-10-12
深圳鼎新融合科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, to provide a test kit and method for genotyping detection, aiming to solve the existing cost of simultaneously detecting the genotypes of four genes of human ADH1B, ALDH2, CYP1A2 and TMPRSS6 High and cumbersome technical issues

Method used

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  • Kit and method for genotyping detection
  • Kit and method for genotyping detection

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Experimental program
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Effect test

Embodiment 1

[0039] A molecular beacon probe method is used for human alcohol metabolism, caffeine metabolism, iron absorption ability-related genotyping detection kit. The kit includes: 1×PCR MIX; positive control; negative control. 1×PCR MIX includes: 1U HS Taq polymerase, 0.5U UNG enzyme, 3.25mM MgCl 2 , 1×PCR Buffer (20mM (NH 4 ) 2 SO 4 , 10mM Tri-HCl, 2mM MgCl 2 ), 200uM dU plus dNTP MIX, the upstream and downstream primer probes of the ADH1B gene, the upstream and downstream primer probes of the ALDH2 gene, the upstream and downstream primer probes of the CYP1A2 gene, and the upstream and downstream primer probes of the TMPRSS6 gene. The specific primer probe sequences are as follows.

[0040] The upstream and downstream primers and molecular beacon probes of the rs1229984 site of the human ADH1B gene are shown in Table 1 below.

[0041] Table 1

[0042]

[0043] The upstream and downstream primers and molecular beacon probes of the rs671 site of the human ALDH2 gene are sh...

Embodiment 2

[0054] Use the kit of the above-mentioned embodiment 1 to carry out genetic detection on human oral cavity samples (assuming that there is only one sample to be tested JYH, sample number JYH).

[0055] Step 1: DNA extraction.

[0056] Step 2: DNA quality detection, the DNA concentration is required to be 5ng / ul-20ng / ul, if the concentration is too low, it can be concentrated, if the concentration is too high, it needs to be diluted to the required range. The value of DNA purity OD260 / OD280 is required to be between 1.7-2.0. Samples that do not meet DNA quality require recollection and DNA extraction.

[0057] Step 3: PCR amplification and sample addition:

[0058] Take out the components of the kit and freeze-thaw; prepare a fluorescence-readable PCR tube.

[0059] Add samples according to the components and volumes in Table 5 below, and add samples to sample JYH, positive control substance, and negative control substance at the same time, shake and mix well after adding th...

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Abstract

The invention belongs to the technical field of molecular biological gene polymorphism detection, and in particular relates to a kit and a method for genotyping detection. The kit consists of a firstprimer pair and a first molecular beacon probe for ADH1B gene rs1229984 site, a second primer pair and a second molecular beacon probe for ALDH2 gene rs671 site, a third primer pair and a third molecular beacon probe for CYP1A2 gene rs762551 site, and a fourth primer pair and a fourth molecular beacon probe for TMPRSS6 gene rs855791 site. With the application of the kit provided by the invention,four genotypes corresponding to the genes and corresponding metabolic absorption types can be rapidly and conveniently detected.

Description

technical field [0001] The invention belongs to the technical field of molecular biology gene polymorphism detection, in particular to a kit and method for genotyping detection. Background technique [0002] Alcohol, that is, ethanol, the metabolic process in the human body mainly occurs in the liver. Ethanol is first converted into acetaldehyde by alcohol dehydrogenase, and acetaldehyde is converted into acetic acid by acetaldehyde dehydrogenase. Acetaldehyde is very harmful to the human body, and it is the chief culprit of alcoholism. It is extremely harmful to various organs in the human body, while acetic acid is basically harmless to the human body. In the process of alcohol metabolism, alcohol dehydrogenase and acetaldehyde dehydrogenase are the main rate-limiting enzymes. The speed of metabolism directly leads to the length of time that acetaldehyde stays in the human body, that is, the degree of harm to the human body. The activity of the speed enzyme varies from pe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12Q1/6883
CPCC12Q1/6858C12Q1/6883C12Q2600/156C12Q2600/16C12Q2531/107C12Q2537/143C12Q2563/107
Inventor 吴宇亮李科铮钟宇萍陈婷李婵杨勇迟海滨
Owner 深圳鼎新融合科技有限公司
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