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Electrochemical method for detecting guanine-containing DNA and counterfeit deterrence authentication method using the same

A detection method and technology of authentication method, applied in biochemical equipment and methods, measurement devices, scientific instruments, etc., can solve the problem of high detection cost, achieve the effect of good selectivity, efficient and rapid separation and enrichment, and reduce detection cost

Inactive Publication Date: 2010-04-14
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for unlabeled electrochemical sensing and detection methods, the detection cost is relatively high

Method used

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  • Electrochemical method for detecting guanine-containing DNA and counterfeit deterrence authentication method using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Commercially available streptavidin-coated magnetic beads (purchased from Promega, with a diameter of about 1 μm) were washed three times with 0.5×SSC buffer according to the requirements, and then combined with biotin-labeled artificially synthesized oligonucleotide chains (capture Probe) mixed (1.25nmol / L capture probe: 1mg magnetic beads), slightly vibrated for 20 minutes, using highly specific binding of avidin and biotin (K d =10 -15 M) DNA probes were immobilized on magnetic particles, magnetically separated and the beads were washed twice with 95 μL TTA (250 mM Tris-HCl, 0.1% Tween 20 and 5% BSA). Add 45 μL of hybridization solution ((750mM NaCl, 150mMSodium citrate) to the magnetic beads, then add the target DNA sequence that can hybridize with the biotin-labeled capture probe, react for 20 minutes, and enrich the target DNA fragments on the magnetic particles Under the action of a certain external magnetic field, the DNA enricher that enriches complementary DN...

Embodiment 2

[0046] Mix the magnetic beads and the capture probe at 1nmol / L capture probe: 1mg magnetic beads, and put the magnetic beads collected above into 0.5mL containing 0.1mol / L MgCl 2 , pH 5.0, 50mM PBS, then add the target DNA sequence that can hybridize with the biotin-labeled capture probe, and then add it to the electrochemical cell, the working electrode is a conductive glass electrode, add Ru(bpy) 3 2+ To its concentration is 1uM, all the same with embodiment 1, its result is as follows figure 1 The dotted line graph is shown.

Embodiment 3

[0048] Mix the magnetic beads and the capture probe at 10 nmol / L capture probe: 1 mg magnetic beads, put the collected magnetic beads into 0.5 mL containing 2 mol / L KCl, pH 9.0, 50 mM PBS, and then add The target DNA sequence to be detected in the hybridization reaction of the capture probe is then added to the electrochemical electrolytic cell, the working electrode is a metal electrode, and Ru(bpy) is added 3 2+ To its concentration is 100uM, all the same with embodiment 1, its result is as figure 1 The dotted line graph is shown.

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Abstract

The invention discloses an electrochemistry DNA method of detecting guanine, which comprises the following steps: 1) preparing or selecting hypoxanthine instead of guanine as capture probe; 2) fixingthe capture probe on the magnetic particle surface, separating magnetically, adding the present sample into the magnetic particle of capture probe. If the present DNA matches the capture probe, it isenriched and separated from samples to the magnetic particle; 3) adding the magnetic particle to the metal inorganic salt buffer solution with the dense of 0.1M at least and pH value ranging from 5 to9 and putting magnetism of electrochemistry electrolytic cell in the electrode working situation; 4) adding Ru (bpy) 32+ to the solution; 5) analyzing the present sample of guanine DNA byelectrochemistry method.

Description

technical field [0001] The invention relates to the sensing and detection technology based on the electrochemical reaction participated by DNA molecule, that is, the electrochemical signal caused by hybridization of DNA molecule can be transmitted and detected through the electrochemical reaction system. Due to the special nature of DNA molecular sequences, this technology is particularly directly related to the application of gene sequences in the field of anti-counterfeiting authentication. Specifically, the present invention relates to an electrochemical method for detecting guanine-containing DNA and an anti-counterfeit authentication method using the same. Background technique [0002] The sensing and detection technology based on the electrochemical reaction in which DNA molecules participate, that is, through the electrochemical reaction system, the electrochemical signals caused by hybridization of DNA molecules can be transmitted and detected, for example, based on ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447C12Q1/68
Inventor 樊春海张治洲
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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