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Method for detecting DNA mutation by nanometer gold

A nano-gold, single-stranded technology, applied in the field of DNA mutation detection, can solve the problem of high cost

Inactive Publication Date: 2010-08-04
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has a high degree of automation and can detect a large number of samples, but requires special instruments and is expensive

Method used

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  • Method for detecting DNA mutation by nanometer gold
  • Method for detecting DNA mutation by nanometer gold
  • Method for detecting DNA mutation by nanometer gold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The detection method is applied to detect the point mutation at codon 245 of the human p53 gene, the base G is mutated to the base A [3] (see literature: Rangel-Lopez A, Maldonado-Rodriguez R, Salcedo-Vargas M, et al.Low density DNAmicroarray for detection of most frequent TP53 missense point mutations.BMC Biotechnol, 2005, 15 (8): 1-13), The specific operation steps are as follows:

[0029] 1) Mix 50 pmol of wild-type single-stranded DNA, an equal amount of probe probe (sequence completely complementary to wild-type single-stranded DNA) and 50 μL of 7.54 nmol / L gold nanoparticles , after standing at room temperature for 10 minutes, add 3 μL of 1mol / L sodium chloride solution, the final concentration of sodium chloride is 0.057, mix well and leave at room temperature for 30 minutes, the obtained system is called system 1.

[0030] 2) Mix 50 pmol of mutant single-stranded DNA, an equal amount of probe probe (sequence completely complementary to wild-type single-stranded...

Embodiment 2

[0035] The detection method is applied to detect the point mutation at codon 245 of the human p53 gene, the base G is mutated to the base A [3] (see literature: Rangel-Lopez A, Maldonado-Rodriguez R, Salcedo-Vargas M, et al.Low density DNAmicroarray for detection of most frequent TP53 missense point mutations.BMC Biotechnol, 2005, 15 (8): 1-13), The specific operation steps are as follows:

[0036] 1) Mix 42 pmol of wild-type single-stranded DNA, an equivalent amount of probe probe (sequence completely complementary to wild-type single-stranded DNA) and 45 μL of 4.81nmol / L gold nanoparticles, After standing at room temperature for 12 minutes, add 4 μL of 1mol / L sodium chloride solution, the final concentration of sodium chloride is 0.082, mix well and leave at room temperature for 33 minutes, the obtained system is called system 1.

[0037] 2) Mix 42 pmol of mutant single-stranded DNA, an equivalent amount of probe probe (sequence completely complementary to wild-type single-...

Embodiment 3

[0042] The detection method is applied to detect the point mutation at codon 245 of the human p53 gene, the base G is mutated to the base A [3] (see literature: Rangel-Lopez A, Maldonado-Rodriguez R, Salcedo-Vargas M, et al.Low density DNAmicroarray for detection of most frequent TP53 missense point mutations.BMC Biotechnol, 2005, 15 (8): 1-13), The specific operation steps are as follows:

[0043] 1) Mix 35 pmol of wild-type single-stranded DNA, an equal amount of probe probe (sequence completely complementary to wild-type single-stranded DNA) and 40 μL of 2.8 nmol / L gold nanoparticles , after standing at room temperature for 9 minutes, add 4 μL of 1mol / L sodium chloride solution, the final concentration of sodium chloride is 0.091, and let it stand at room temperature for 35 minutes. The obtained system is called system 1.

[0044] 2) Put 35 pmol of mutant single-stranded DNA, an equivalent amount of probe probe (completely complementary sequence to wild-type single-strande...

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Abstract

The invention discloses a DNA mutation test method in use of nano-Au, relates to a DNA mutation test method, and provides a DNA mutation test method that uses a nano-Au probe. The DNA mutation test method leads wild single-stranded DNA, a probe that is completely complementary with a wild single-stranded DNA sequence and the nano-Au to be mixed, statically placed, added with a sodium chloride solution, and then statically placed, thus obtaining an architecture 1; the DNA mutation test method further leads mutant single-stranded DNA, the probe that is completely complementary with the wild single-stranded DNA sequence and the nano-Au to be mixed, statically placed, added with the sodium chloride solution, and then statically placed, thus obtaining an architecture 2; the sodium chloride solution is added into and evenly mixed with the architecture 1, the ultraviolet-visible absorption spectrum detection is implemented, and an absorbance value obtained at a 520nm position is taken as an absorbance value 1; the sodium chloride solution is added into and evenly mixed with the architecture 2, the ultraviolet-visible absorption spectrum detection is implemented, and an absorbance value obtained at a 520nm position is taken as an absorbance value 2; when the absorbance values 2 and 1 have marked difference, mutation is determined to exist. The DNA mutation test method does not need any modification of the DNA and the nano-Au, thus simplifying sample processing.

Description

technical field [0001] The invention relates to a method for detecting DNA mutations, in particular to a method for detecting DNA mutations with nanometer gold probes. Background technique [0002] Gene mutation refers to the change of base pair composition or sequence in the structure and function of genomic DNA molecules, mainly including base substitution, deletion or insertion. Since gene mutations are closely related to the occurrence of various genetic diseases and tumors, the detection of gene mutations is of great significance for the early detection and diagnosis of genetic diseases or tumors. [0003] The methods for detecting gene mutation mainly include gel electrophoresis, DNA sequencing and DNA chip technology. The basic principle of gel electrophoresis is: DNA with different conformations has different mobility in the gel, and whether there is a gene mutation can be judged according to the change of the mobility. Although this method is simple, its sensitivi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/33C12Q1/68
Inventor 孙莉萍李辉张建锋张召武王霜张其清
Owner XIAMEN UNIV
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