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Human ALDH2 genotype detection kit

A detection kit and genotype technology, applied in the field of genetic engineering, can solve the problems of low detection sensitivity, complicated operation, cumbersome operation, etc.

Active Publication Date: 2014-10-29
JIANGSU WEIHE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest advantage of the gene chip method is high throughput, but there are also many disadvantages, including high technical cost, complicated operation, low detection sensitivity and poor repeatability, etc.
PCR-RFLP, that is, PCR-restriction fragment length polymorphism technology, has also been applied to the detection of ALDH2 gene, but it has not been popularized due to the disadvantages of cumbersome operation, time-consuming, and low reliability of results.

Method used

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  • Human ALDH2 genotype detection kit
  • Human ALDH2 genotype detection kit
  • Human ALDH2 genotype detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1 Raw materials and equipment:

[0027] 1.1 Contents of the kit:

[0028] 1) 8-well 200μl PCR reaction plate, detect one sample per 2 wells

[0029] 2) 2 pairs of specific upstream and downstream primers for different subtypes of ALDH2, in each 2 wells, add primer A to the first well, add primer B to the second well, and add a pair of internal control primers to each well , all primers are lyophilized at the bottom of the PCR reaction plate.

[0030] The nucleotide sequences of primer A and primer B are as follows:

[0031]

[0032] Internal control primers, the nucleotide sequence is as follows:

[0033] 5’-TGCATCTGGACATGCTTGCT-3’

5'-TGGCTGGAGGAGACTCCAAA-3'

[0034] 3) PCR reaction system

[0035] DNA polymerase: Polyethylene glycol (PEG)-modified hot-start Taq polymerase that can tolerate PCR reaction inhibitors in blood;

[0036] Reaction buffer: composition and each component content are as follows: 0.18mM deoxynucleotide (dNTP), 1.8m...

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Abstract

The invention provides a human ALDH2 genotype detection kit comprising a PCR amplification primer, DNA polymerase and a reaction buffer solution; wherein the DNA polymerase is a polyethylene glycol (PEG) modified hot-start Taq polymerase which can tolerate the PCR reaction inhibitors in blood; and further provides the nucleotide sequence of the PCR amplification premier. The kit has the following advantages: (1) the DNA polymerase is a polyethylene glycol (PEG) modified hot-start Taq polymerase which can tolerate the PCR reaction inhibitors in blood, thus can realize the human whole-blood PCR amplification without carrying out nucleic acid extraction, and is capable of directly carrying out PCR amplification in blood; (2) the kit can carry out detection on a sample ALDH2 genotype.

Description

technical field [0001] The invention relates to a human ALDH2 genotype detection kit, which belongs to the technical field of genetic engineering. Background technique [0002] Aldehyde dehydrogenase (ALDH) is a tetrameric protein that catalyzes the oxidation of acetaldehyde and other aliphatic aldehydes. In human organs and tissues, there are 19 isoenzymes of ALDH, the common ones are ALDH1~ALDH4, of which ALDH2 is located in the mitochondria, while ALDH1, ALDH3 and ALDH4 are located in the cytosol. Based on substrate specificity and subunit composition, only ALDH1 and ALDH2 are considered to be the two major isoenzymes in the human liver, and only ALDH2 exhibits genetic polymorphisms. [0003] The ALDH2 gene is located on human chromosome 12, and nearly a hundred SNP sites have been discovered so far. The main polymorphism is the rs671 site, that is, the ALDH2 gene has a point mutation (Glu487Lys) at exon 12, which makes the catalytically active wild The type allele ALDH...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2521/101C12Q2527/125
Inventor 陈炤源王浩
Owner JIANGSU WEIHE BIOTECH
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