Nanometer detecting probe chip without amplifying genom DNA and detection method

A technology of nano-probe and detection method, which is applied in the field of nano-probe chip to achieve the effects of convenient popularization, good repeatability and simple operation

Active Publication Date: 2008-05-21
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Application Information

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Problems solved by technology

[0009] The technical problem to be solved in the present invention is to provide a nano-chip and detection method without gene amplification for the shortcomings of current PCR and traditional chip fluorescence detection methods

Method used

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  • Nanometer detecting probe chip without amplifying genom DNA and detection method

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Embodiment Construction

[0036] The present invention will be further described below in conjunction with the embodiments of the present invention and the accompanying drawings.

[0037] Example Nanoprobe chip for P53 gene detection

[0038] 1. Prepare materials and reagents

[0039] (1) Chip: self-made;

[0040] (2) EasyHtyb hybridization solution: purchased from Roche Company;

[0041] (3) All probes were synthesized in Dalian Bao Biological Engineering (Dalian) Co., Ltd. (Dalian TaKaRa Company)

[0042] (4) 15nm and 30nm gold nanoparticles were purchased from Shanghai Water Source Biotechnology Co., Ltd.

[0043] (5)NaNO 3 Washing solution: 1M NaNO 3 300ml

[0044] 0.2M Na 2 HPO 4 30.5ml

[0045] 0.2M NaH 2 PO 4 9.5ml

[0046] Dilute to 1000ml

[0047] (6) PB solution: 0.3mol / L NaNO 3 and 10mmol / L Na 2 HPO 4 / NaH 2 PO 4 , PH7.0

[0048] (7) Silver stain enhancement solution: 60 μl of 1% gelatin

[0049] 2.55g citri...

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Abstract

The invention relates to a nanosized probe chip which does not need to magnify the genome DNA and a detection method; the invention is characterized in that the detection of nucleic acid with unmarked target produces high-sensibility identification signal through two steps of continuous sandwich crossbreed reaction with the capture probes which are equipotentially and specialally fixed on the surface of the chip and special oligonucleotide probes of the marked nanosized gold with the magnification effect of the nanosized gold signal which is enhanced by silver stein; and then microscope is used directly for observation or ordinary optical scanner is used for scanning to directly obtain the corresponding crossbreed results; the detection report can be obtained through the analysis of relevant software. Compared with the traditional method based on fluorescent signal detection, the method greatly increases the sensibility and specials of the detection and can detect the targeted gene and single base group mutation of unmagnified genome DNA sample.

Description

technical field [0001] The invention relates to a nano-probe chip and a detection method without amplification of genomic DNA and the nano-probe chip detection method, in particular to a nano-probe chip detection method for directly detecting target genes and gene mutations in genomic DNA. The invention belongs to the field of nano probe chip. Background technique [0002] The study of gene mutation has become one of the hot spots in life science research. Detection methods have also developed rapidly. For the detection of gene mutations, Southern blotting was mainly used 20 years ago, which can screen out mutations such as gene deletions, insertions, and frameshift recombination. However, this method cannot detect single nucleotide polymorphism (SNP) point mutations, and complex DNA sequencing analysis methods are mainly used for point mutations. Polymerase chain reaction (polymerase chain reaction, PCR) technology is the most significant progress in mutation research, w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 毛红菊赵建龙贾春平金庆辉
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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