GSTT1 genotyping method based on quantitative PCR technology

A detection method and technology, which is applied in the field of GSTT1 typing detection based on quantitative PCR technology, can solve problems affecting the accuracy of gene trait analysis, difficult heterozygous genotypes, difficult experiment reliability, etc., and achieve simplified detection and result analysis Process, avoidance of contamination, effect of prevention of PCR product contamination

Inactive Publication Date: 2017-09-15
上海龙鼎医药科技有限公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it is simple, it is easy to cause laboratory pollution because the electrophoresis operation must open the tube to add samples and electrophoresis, and it is difficult to ensure the reliability of the exper

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
  • GSTT1 genotyping method based on quantitative PCR technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] (1) Preparation of normal human genomic DNA for testing: take normal human oral and throat swabs, extract and prepare with Chlex100, and dilute the concentration of normal human genomic DNA to 10 ng / μl;

[0039] (2) Design and synthesis of PCR-specific primers and probes: Design and synthesize gene-specific primers SEQ1, SEQ2 and GSTT1-specific TAQman gene probe marker Fam according to the human GSTT1 gene sequence; design an internal reference gene-specific PCR primer SEQ4 based on the human ALbumin gene , SEQ5 and probe SEQ6 mark Vic;

[0040] (3) GSTT1 quantitative PCR detection:

[0041] 1) Primer-probe mix configuration:

[0042] 2) Reaction system: 1.5 μl of primer-probe mixture, 12.5 μl of 2-fold TIANtoughGenotypingqPCCRPreMix (Probe), 1.5 μl of standard template, and 9.5 μl of ddH2O.

[0043] 3) Quantitative PCR instrument: ABI7500

[0044] 4) Reaction conditions: 95°C, 2min; 95°C, 15s---60°C, 32s, 42 cycles.

[0045] (4) Gene verification result read:

[0...

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 GSTT1 genotyping method based on quantitative PCR technology. According to the GSTT1 genotyping method based on quantitative PCR technology, GSTT1 gene specific primers SEQ1 and SEQ2 are used for amplification of GSTT1 gene segments, and GSTT1 gene specific TAQman probe SEQ3-Fam is designed in amplification regions defined by the GSTT1 gene specific primers at the same time; ALbumin specific PCR primers SEQ4 and SEQ5 are used for amplification of ALbumin gene segments, and ALbumin gene specific TAQman probe SEQ6-Vic is designed in amplification regions defined by the ALbumin gene specific primers at the same time. The GSTT1 genotyping method is capable of protecting PCR products from being polluted, avoiding pollution caused by toxic DNA coloring agents, simplifying detection and result analysis process greatly, and increasing detection flux.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a GSTT1 typing detection method based on quantitative PCR technology. Background technique [0002] GSTT1 is an important metabolic enzyme involved in the transformation and metabolism of endogenous and exogenous substances, endowing metabolites with new biological properties. GSTT1 is polymorphic in humans, and GSTT1 deletion is a common genotype. There are differences in the transformation function of the individual geomaterials of different GSTT1 genotypes. GSTT1 genotyping is used in the study of various biological traits. Traditionally, since the GSTT1 deletion polymorphism can be detected by ordinary PCR, conventional gene-specific PCR and agarose electrophoresis detection methods are often used to confirm the GSTT1 genotype. Although it is simple, it is easy to cause laboratory pollution because the electrophoresis operation must be opened to add samples and elec...

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): C12Q1/68C12Q1/48
CPCC12Q1/6858C12Q2531/113C12Q2561/101
Inventor 杨随娟杨燕燕刘虎
Owner 上海龙鼎医药科技有限公司
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