Gene chip for integrated detection and on-substrate amplification method

A gene chip and gene amplification technology, applied in the field of gene chips, can solve the problems of easy amplification of pollution, false positives, cross-contamination, etc., and achieve the effect of solving the mutual interference of primers, simple and universal instruments, and improving efficiency.

Inactive Publication Date: 2014-08-06
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, this technology has the following problems: first, because of the high sensitivity of PCR amplification, cross-contamination is easy to occur in the process of amplification and transfer, and the contamination is easy to be amplified, and the phenomenon of "false positive" occurs, which not only affects the The judgment of the results often needs to be verified by other experiments.
However, it still has the step of DNA hybridization, for which it also specially designs and transforms the amplification tube.

Method used

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  • Gene chip for integrated detection and on-substrate amplification method

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Embodiment 1

[0031] The gene chip for integrated detection and the on-chip amplification method of the present invention is to directly put the gene chip made by fixing specific nucleic acid primers on the substrate into a common amplification tube for PCR amplification. The special DNA sequence immobilized on the substrate is put into the amplification tube as a part of the primer, and the corresponding PCR amplification system is added, including the required primers with fluorescent labels, the gene to be detected, and the amplification system (dNTP, corresponding Enzymes and buffers, etc.) for PCR amplification. After the gene amplification is completed, the fluorescent signal on the substrate is detected to obtain gene information. The amplification system in the amplification tube includes a substrate for on-chip PCR reaction in the tube, a primer system immobilized on the substrate, a labeled nucleic acid primer system, and an amplification system (dNTP, corresponding enzymes and buf...

Embodiment 2

[0040]The gene chip and on-chip amplification method for integrated detection of the present invention, the gene chip made by fixing specific nucleic acid primers on the substrate, is directly put into an ordinary amplification tube for PCR amplification, and is used for detecting GM soy. On the glass chip in the tube, the specific primers of the transgenic soybean transfer gene are immobilized by the spotting instrument; the labeled specific primers, the gene to be tested, the amplification system (dNTP, corresponding enzyme and buffer, etc.) are added to the amplification tube. ). After PCR amplification, through fluorescence scanning, if fluorescence appears, the gene amplification reaction occurs, and the gene to be tested contains the transferred gene, indicating that the sample comes from transgenic soybeans.

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Abstract

The invention discloses a gene chip for integrated detection and an on-substrate amplification method. The on-substrate amplification method comprises the following steps: (1) fixing specific nucleic acid primers on the substrate; (2) placing the substrate, on which the specific nucleic acid primers are fixed, in an amplification tube; (3) adding prepared a gene extract to be detected and an amplification system to the amplification tube; (4) placing the amplification tube in a gene amplification instrument for amplification; (5) after completion of the gene amplification, detecting fluorescent signals on the substrate so as to acquire genetic information. In the on-substrate amplification method, the general step of nucleic acid hybridization is removed, the benefits are greatly increased, and the cost is reduced; the specific nucleic acid primers are fixed on the substrate, and then the specific nucleic acid primers and the substrate are placed in the amplification tube for on-substrate PCRs (Polymerase Chain Reactions), so that the PCR reactions can be realized separately and the problem that primers interfere with one another during multiple PCRs can be solved; multiple pieces of genetic information can be detected integrally at the same time, and contamination, which can be generated in the amplification process and the product transfer process, is avoided; the used instrument is simple and universal, so that a common PCR instrument and a fluorescence microscope can complete the detection.

Description

technical field [0001] The invention relates to an on-chip PCR amplified gene chip, which can be used for integrated detection of gene information, so that corresponding functions such as gene amplification and detection can be completed in the same closed space without transfer operation. Background technique [0002] At present, polymerase chain reaction (PCR) is one of the indispensable technologies in various detection and research processes involving nucleic acid molecules such as molecular biology, gene detection, disease detection and diagnosis. The principle of PCR is the semi-conservative replication of DNA (deoxyribonucleic acid, Deoxyribonucleic acid). Double-stranded DNA can be denatured and unwound into single strands under the action of various enzymes. With the participation of DNA polymerase, according to the principle of complementary base pairing Replicate into two identical molecular copies. DNA can also be denatured and melted at high temperature, and ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C40B40/06
CPCC12Q1/686C12Q2565/501
Inventor 吴凌伟秦煊
Owner NANCHANG UNIV
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