Method for detecting esophagus cancer susceptibility gene PTEN polymorphism with Hae III

A technology for susceptibility genes and esophageal cancer, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve problems such as small scope of application, high detection cost, and complicated operation, and the method is simple and easy Good, easy to recognize, low cost effect

Inactive Publication Date: 2017-02-22
ZHENGZHOU UNIV
View PDF0 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims to solve the existing polymorphism detection method of esophageal cancer susceptibility gene, and overcome the problems of comp...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for detecting esophagus cancer susceptibility gene PTEN polymorphism with Hae III
  • Method for detecting esophagus cancer susceptibility gene PTEN polymorphism with Hae III
  • Method for detecting esophagus cancer susceptibility gene PTEN polymorphism with Hae III

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1. Determination of Human PTEN rs701848 Polymorphism in Human Esophageal Cancer Tissue Specimens

[0071] In a specific embodiment of the present invention, the specific steps for detecting the human PTEN gene polymorphism rs701848 are as follows:

[0072] (l) Obtain surgically removed esophageal cancer tissue, and use the phenol-chloroform method to extract the genomic DNA of the esophageal cancer tissue as the DNA to be tested. The extraction steps are as follows:

[0073] 1) Thaw the esophageal cancer tissue block, wash away blood stains with normal saline, cut 0.1g of tissue for grinding, add 1ml of sterilized water, mix by inverting, centrifuge at 10000rpm for 10min, discard the supernatant, repeat the above steps twice .

[0074] 2) Add 200 μl of DNA lysate and 5 μl of proteinase K, mix well, and digest overnight in a water bath at 55°C.

[0075] 3) After the digestion is completed, add an equal volume of phenol-chloroform mixture (1:1) and shake vigorou...

Embodiment 2

[0085] Example 2. Determination of human PTEN rs701848 polymorphism in human peripheral blood whole blood samples

[0086] The steps are basically the same as in Example 1, except that the following method is used to extract genomic DNA from human peripheral blood as the DNA to be tested.

[0087] Follow the operation steps of NEP004-1 Whole Blood Genomic DNA Extraction Kit to extract the genomic DNA of the blood sample to be tested. The specific steps are as follows:

[0088] 1) After adding 300 μl of blood cells into a 1.5ml centrifuge tube, add 900 μl of cell lysate and mix evenly. After placing it on ice for 10 minutes, centrifuge at 12,000 rpm for 1 minute in a centrifuge, discard the supernatant, add 900 μl of cell lysate again, and use a gun to After blowing up the precipitate and mixing it, repeat the above steps;

[0089] 2) Add 600 μl of solution B solution to the precipitate, gently blow up the precipitate with a pipette gun, add 10 μl of proteinase K and mix well,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for detecting esophagus cancer susceptibility gene PTEN polymorphism with Hae III. Primer sequences are shown as SEQ ID NO:1 and SEQ ID NO:2. The method includes the steps that genomic DNA of a sample is extracted; a forward primer and a reverse primer for amplifying sequences near the human PTEN gene polymorphism rs701848 site are provided, PCR amplification is conducted with the extracted genomic DNA to be detected as the template, and an amplification product is obtained; the obtained amplification product is subjected to digestion with the restriction enzyme Hae III, and a corresponding digestion product is obtained; the digestion product is subjected to electrophoresis with 3% agarose gel so as to judge the genotypes of PTEN gene polymorphism rs701848. The method is simple, easy to implement, rapid, efficient and low in cost, the digestion result is easy to identify, and a simple way is provided for genotyping of esophagus cancer susceptibility gene PTEN polymorphism rs701848.

Description

technical field [0001] The invention belongs to the technical field of gene polymorphism detection, in particular to a HaeIII method for detecting the polymorphism of esophageal cancer susceptibility gene PTEN. Background technique [0002] At present, single nucleotide polymorphism (single nucleotide polymorphism, SNP) is the result of environmental selection in the long-term evolution of human beings, and has become the marker of the third generation of molecular inheritance, with the advantages of high marker density, stability, and easy typing and detection , is one of the genetic basis of differences between races and individuals, has been identified as the basis of susceptibility to a variety of genetically related diseases, and has become an evaluation index for the degree of disease risk. PCR-RFLP (Polymerase Chain Reaction restriction fragment length polymorphisms) is an in vitro enzymatic synthesis of specific nucleotide fragment technology that simulates the in vi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6886C12Q2600/156C12Q2531/113C12Q2521/301
Inventor 代丽萍谢伟张有改段富交李记天王凯娟宋春花张建营
Owner ZHENGZHOU UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap