Method for detecting mutation of gene point through combination of amplification of equipmential specificity and Nano metal probe

A nano-gold probe and allele-specific technology, applied in the field of gene point mutation detection, can solve the problems of complicated operation and time-consuming, and achieve the effects of intuitive results, simple operation and low detection cost.

Inactive Publication Date: 2008-06-18
SOUTH CHINA NORMAL UNIVERSITY
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these traditional detection methods based on electrophoretic analysis have the disadvantages of complex operation, time-consuming, and the need to use radioactive elements or harmful substances such as ethidium bromide

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0010] We applied the gene point mutation detection method combined with allele-specific amplification and nano-gold probes to detect the mutation hotspot at codon 12 of the K-ras gene. The specific steps are as follows:

[0011] (1) Design mutant primers according to the K-ras gene sequence;

[0012] (2) using wild-type and mutant gene sequences as templates to perform allele-specific amplification;

[0013] (4) Add 10 μl of the above amplification product to 200 μl of nano-gold sol;

[0014] (5) Add 60 μl of PBS solution (10 mM, 0.2 M NaCl), observe the color change, and take pictures of the experimental results with a digital camera.

[0015] The results showed that the color of the mutant gene changed from red to blue; the wild-type gene remained red.

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

A method for deteting the mutation at gene site by combining allelic specific amplification with nano-gold probe inclues designing the allelic specific primer complementary with mutant gene but not with wild gene, allelic specific amplifying to obtain double-stranded DNA (product A) and residual single-stranded DNA-primer (product B), and sequentially adding said product and electrolyte solution to nano-gold sol. The color of solution is changed from red to blue for the product A or its not changed for the product B.

Description

technical field [0001] The invention relates to a gene point mutation detection method for gene point mutation detection and single nucleotide polymorphism analysis, which combines allele-specific amplification and nanometer gold probes. Background technique [0002] At present, common methods for detecting gene point mutations include: restriction fragment length polymorphism, denaturing gradient gel electrophoresis, chemical mismatch cleavage, enzymatic mismatch cleavage, single-strand conformational polymorphism, allele-specific probe hybridization, Ligase chain reaction and competitive oligonucleotide primers, etc. Most of these traditional detection methods based on electrophoretic analysis have the disadvantages of complex operation, time-consuming, and the need to use radioactive elements or harmful substances such as ethidium bromide. Contents of the invention [0003] The object of the present invention is to provide a gene point mutation detection method for gen...

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 Patents(China)
IPC IPC(8): C12Q1/68
Inventor 邢达周政朱德斌
Owner SOUTH CHINA NORMAL UNIVERSITY
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