Supercharge Your Innovation With Domain-Expert AI Agents!

Technology for detecting polymorphism of human TERF1 gene rs3863242 site

A site polymorphism and gene technology, which is applied in the field of single nucleotide polymorphism determination to achieve good yield and specificity, and reduce the cost of determination.

Inactive Publication Date: 2016-02-03
郑州市职业病防治院
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no restriction endonuclease has been found that can recognize the base sequence containing the polymorphic site of TERF1 gene rs3863242, and PCR-RFLP technology cannot be used for detection, and new detection technology needs to be developed

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
  • Technology for detecting polymorphism of human TERF1 gene rs3863242 site
  • Technology for detecting polymorphism of human TERF1 gene rs3863242 site
  • Technology for detecting polymorphism of human TERF1 gene rs3863242 site

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Extraction of Human Peripheral Blood Leukocyte DNA Determination of rs3863242 Polymorphism

[0059] 1 Materials and methods

[0060] 1.1 Main reagents and instruments

[0061] Reagent: 2×PCRMix (including 4 kinds of dNTP mixture, Mg 2+ , TaqDNApolymerase, TaqAntibody, buffer, DMSO and ammonium sulfate), restriction endonuclease HpyCH4IV (NEB Company), agarose (Biowest Company), and primers were synthesized by Shanghai Sangon Company.

[0062] Instruments: Model 9600 PCR instrument (PE Company), mini electrophoresis tank (PharmaciaBiotech, EPS1000), GelDoc2000 gel imager (Bio-RAD Company).

[0063] 1.2 Extract DNA from peripheral blood leukocytes as the template of genomic DNA to be tested

[0064] EDTA-K 2 Collect 1mL of human peripheral blood with an anticoagulant tube, and separate leukocytes by referring to the leukocyte separation method in "Practical Flow Cytometry Color Atlas", and extract leukocyte genomic DNA by referring to the sodium chloride sal...

Embodiment 2

[0088] Example 2 Determination of the polymorphism of TERF1 gene rs3863242 in whole blood samples of human peripheral blood

[0089] The main reagents and instruments are the same as in Example 1; refer to the phenol-chloroform extraction method in "Molecular Cloning Technology" to extract peripheral blood genomic DNA, and use it as a human genomic DNA template to be tested.

[0090] The sequence search is the same as in Example 1, the upstream primer used in the PCR reaction is SEQNO:8, and the downstream primer is SEQNO:9.

[0091] Upstream primer: 5'AATTCAGTTGACATGTAGCACTGCCA3' (SEQ ID NO: 8); Downstream primer: 5'TAAACATTTCTTAGATTTTTAAAAGATTTTACATTTA A C3' (SEQ ID NO: 9).

[0092] Take genomic DNA (50-80ng), upstream and downstream primers (final concentration: 0.1-1 μM), 2×PCRMix 10 μL ((including 4 kinds of dNTP mixture, Mg 2+ , TaqDNApolymerase, TaqAntibody, buffer, DMSO and ammonium sulfate) and sterilized double distilled water to form a 20 μL reaction system, and a...

Embodiment 3

[0107] Example 3 Determination of rs3863242 polymorphism of TERF1 gene in human peripheral blood clot specimen

[0108] The main reagents and instruments are the same as in Example 1; refer to the phenol-chloroform extraction method in "Molecular Cloning Technology" to extract the genomic DNA of the peripheral blood clot, and use it as the human genomic DNA template to be tested.

[0109] The upstream primer used in the PCR reaction is SEQNO:13, and the downstream primer is SEQNO:14.

[0110] Upstream primer: 5'CAAATTCAGTTGACATGTAGCACTGCCA3' (SEQ ID NO: 13);

[0111] Downstream primer: 5'GACTTCTCTAAACATTTCTTAGATTTTTAAAAGATTTTACATTTA A C3' (SEQ ID NO: 14).

[0112] Except that the annealing temperature of PCR amplification is 64°C, the other reaction conditions are the same as in Example 1; the enzyme digestion reaction is the same as in Example 1, and the enzyme digestion product is concentrated by 1 times, and electrophoresis is carried out under the condition of 2.5% agaro...

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 technology for detecting polymorphism of a human TERF1 gene rs3863242 site. The method comprises the following steps: extracting to-be-detected human genome DNA; providing an upstream primer and a downstream primer of a nearby sequence for amplifying the human TERF1 gene rs3863242 site, wherein the downstream primer has a mismatched base A; implementing a PCR amplification reaction by virtue of the upstream primer and the downstream primer by taking the to-be-detected genome DNA as a template so as to obtain an amplification product containing an AYGT segment, wherein Y (T or C) is a complementary base of an undetermined base R (A or G) on the human TERF1 gene rs3863242 site; providing a restriction enzyme; digesting the amplification product by virtue of the restriction enzyme so as to obtain a corresponding enzyme-digested product; and determining whether the undetermined base R on the human TERF1 gene rs3863242 site is A or G according to the enzyme-digested product. The determination means disclosed by the invention is not only rapid and reliable, but also capable of greatly reducing a determination cost.

Description

technical field [0001] The present invention relates to methods for determining single nucleotide polymorphisms. Background technique [0002] SNP (Single Nucleotide Polymorphism) refers to the DNA sequence variation caused by the change of a single nucleotide, including single base substitution, insertion and deletion. Gene polymorphism is an individual's innate genetic characteristics, does not change with the environment, and is not a specific or non-specific clinical manifestation of a certain disease, so its application does not have diagnostic and therapeutic functions. [0003] Gene polymorphism detection is widely used in the field of genetics and medicine, examples are as follows: 1. To study genetic phenomena such as the origin and evolution of species. According to the genetic variation, the information of biological macromolecules can be obtained, the history of biological evolution can be inferred, and the evolution relationship of species can be clarified. It...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2521/301C12Q2525/117
Inventor 张育红李磊张辉李露李海娟王刚刘伟俊刘素香李志凯
Owner 郑州市职业病防治院
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com