Human nucleotide repair protein gene main polymorphism site fast detection method and kit thereof

A polymorphic site and protein repair technology, applied in the field of molecular biology, can solve the problems of time-consuming and cumbersome operation, poor specificity and sensitivity, time-consuming and expensive sequencing and chip technology, etc.

Inactive Publication Date: 2008-05-28
吕成伟
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] (4) Economical and practical. Traditional SNP or point mutation detection methods, such as PCR-RFLP, PCR-SSCP, PCR-SSO, etc., mostly involve post-processing such as electrophoresis, enzyme digestion, color development, and photography. The operation is time-consuming and cumbersome, and the specificity and The sensitivity is poor, it cannot be automated and high-throughput detection, and it is easy to cause cross-contamination. Sequencing and chip technology are time-consuming and expensive

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
  • Human nucleotide repair protein gene main polymorphism site fast detection method and kit thereof
  • Human nucleotide repair protein gene main polymorphism site fast detection method and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0033] The composition of the kit of the present invention is as follows:

[0034] Reaction mixture: 1×PCR buffer (0.1% TritonX-100; 2.0~6.0mmol / L MgCl 2 ; 8mmol / L(NH 4 ) 2 SO 4 ; 50mmol / L KCl; 10mmol / LTris-HCl(pH8.0)); contains two primers each 0.3μmol / L, Probe-wt0.2μmol / L, Probe-mu0.3μmol / L; dNTP each 250μmol / L, HotTaq enzyme 1.0U, total reaction volume 50μl.

[0035] Sample extraction solution: (1). Absolute ethanol; (2). 1.5% triethanolamine lauric acid sulfate (TLS); (3). Chloroform: isoamyl alcohol (24:1); (4). TE (pH 8 .0).

[0036] R194W mutant wild-type probe sequence:

[0037] Probe-wt: FAM-5’-GCGCTCAGCCGGATCAACAAGACAGCGC-3’-DABCYL

[0038] R194W mutant mutant probe sequence:

[0039] Probe-mu: HEX-5’-GCACGGCTCTCTTCTTCAGCTGGATCGTGC-3’-DABCYL

[0040] R280H mutant wild-type probe sequence:

[0041] Probe-wt: FAM-5’-CGCTGCCAGCTCCAACTCGTACCAGCG-3’-DABCYL

[0042] R280H mutant mutant probe sequence:

[0043] Probe-mu: HEX-5’-CGCTGCCAGCTCCAACTCATACCAGCG-3’-DABCYL

[0044] R399Q...

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 rapid testing method for main polymorphism site of repair protein gene (XRCC1) of human ribonucleotide. According to designed primers of genetic DNA sequence of human chromosome XRCC1, the invention employs polymerase chain reaction (PCR) specific technique to amplify a fragment of DNA sequence respectively including mutation sites of R194W, R280H and R399Q, respectively uses a pair of molecular fluorescent probes to be hybridized with amplified product. Whether a sample includes mutation of R194W, R280H and R399Q is tested by real-time detection of a fluorescent PCR instrument to corresponding specific fluorescent intensity alternation of reaction tubes or by direct detection of specific fluorescent intensity of reaction tubes via a fluorescence spectrophotometer after PCR. The method is convenient and rapid, high in specificity, economic and practical, which is applicable for rapid detection of large-scaled samples. The invention further provides kits applicable for clinical application.

Description

Technical field [0001] The invention belongs to the field of molecular biology. Background technique [0002] Lung cancer is the number one cancer and has become a major cause of cancer death. Platinum drugs that play an anti-tumor effect with DNA damage as the main mechanism mainly include cisplatin (DDP), carboplatin (CBP) and oxaliplatin (L-OHP). They are currently the basic chemotherapeutic drugs used to treat advanced lung cancer, but The sensitivity of different individuals to platinum drugs varies greatly, which affects the clinical efficacy of such drugs. Studies have shown that if tumor cells have strong DNA repair capabilities, the efficacy of these drugs is poor, and vice versa. [0003] The XRCC1 gene is an important component in the base excision repair / single-strand break repair system. Its encoded product interacts with DNA ligase III to repair DNA single-strand breaks and performs base excision repair together with DNA polymerase β to maintain Genome stability is ...

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
Inventor 吕成伟
Owner 吕成伟
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