Membrane transfer method for testing gene chip

A technology of gene chip and detection method, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., and can solve the problems of expensive chip detection equipment and expensive chips

Inactive Publication Date: 2002-01-09
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Abstract
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  • Claims
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Problems solved by technology

Thus improving the current situation that chips are expensive and chip testing equipment is expensive and difficult to promote

Method used

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  • Membrane transfer method for testing gene chip
  • Membrane transfer method for testing gene chip
  • Membrane transfer method for testing gene chip

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Embodiment Construction

[0023] The present invention will be further described below in conjunction with the embodiments of the present invention and the accompanying drawings.

[0024] The concrete method of the membrane transfer detection method of gene chip of the present invention is as follows:

[0025] 1. Prepare materials and reagents

[0026] ① DNA chip: self-made;

[0027] ②PCR mixture: 200 μM dATP, 200 μM dGTP, 200 μM dCTP, 200 μM dTTP, 1.25 U pfu DNA polymerase, 0.2 μM upstream and downstream primers each, and the 5′ ends of the primers were labeled with digoxigenin or biotin. If the primers are not labeled with digoxin or biotin, change the concentration of dTTP to 80 μM, and add 40 μM digoxin or biotin-labeled dUTP;

[0028] ③EasyHtyb: purchased from Roche;

[0029] ④ lotion I: 150mM NaCl, 15mM sodium citrate, 0.1% sodium dodecyl sulfate (SDS);

[0030] ⑤ Lotion II: 100mM Maleic acid, 150mM NaCl, 0.3% (v / v) Tween22, pH7.5;

[0031] ⑥Antibody: 1U / ml Anti-Digoxigenin-Ap or Anti-Biotin...

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Abstract

A membrane transfer method for testing gene chip includes such steps as adding digoxin or biotin label in the PCR amplification process of the specimen to be tested, hybridizing the PCR resultant with chip, binding antibody with digoxin or biotin label, and transferring the hybridized result to nylon or cellulose ucetate membrane wetted by colouring developing liquid for higher intensity of hybridized signals and higher test sensitivity.

Description

technical field [0001] The invention relates to a gene chip, in particular to a membrane transfer detection method of the gene chip. Background technique [0002] Gene chips are formed by using the intensive and parallel processing principles of microelectronic chip technology to orderly solidify a large number of biological samples with characteristic sequences of biological significance on silicon substrates or glass. Gene chips can be used to quickly obtain or process A large amount of life information (including gene identification, gene mutation detection, gene expression profile detection, etc.), which has been successfully applied in the fields of gene expression, gene mutation, gene polymorphism research and gene diagnosis. [0003] When using a gene chip, the target sample nucleic acid often needs to be amplified or cloned or reverse transcribed (mRNA) by polymerase chain reaction (PCR, hereinafter referred to as PCR), and isotope or fluorescent labeling is carried ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 赵建龙景奉香孙悦张为毛红菊赵辉
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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