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NAT2 gene polymorphism fluorescence PCR melting curve detection kit

A technology of gene polymorphism and detection kit, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of high cost of gene chip technology, long detection cycle of sequencing method, contamination of false positive results, etc. Achieve the effects of avoiding repeated freezing and thawing, short detection time, and easy results

Inactive Publication Date: 2017-02-01
XIAMEN ZEESAN BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Inhibitory fragment length polymorphism analysis requires enzyme digestion and electrophoresis of PCR products, and the operation steps are cumbersome and easy to cause pollution
The cost of gene chip technology is high, the operation is complicated, and the results will also have different degrees of false positives
The throughput of the ARMS-PCR method is very limited, and PCR post-processing is required, which is prone to contamination and false positive results
Sequencing method has a long detection cycle and is not suitable for rapid analysis of clinical samples

Method used

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  • NAT2 gene polymorphism fluorescence PCR melting curve detection kit
  • NAT2 gene polymorphism fluorescence PCR melting curve detection kit
  • NAT2 gene polymorphism fluorescence PCR melting curve detection kit

Examples

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Embodiment 1

[0035] A NAT2 gene polymorphism fluorescence PCR melting curve detection kit, which is a dry kit (no solvent exists, each reagent and nucleotide is dry powder), including a first probe, a second probe, a A third probe, a fourth probe, a forward primer and a reverse primer, specifically, the single-tube reaction system includes: 1×PCR buffer, 2U Taq DNA polymerase, 200μM dNTP, 4.5mM MgCl 2 , forward primer 4pmol, reverse primer 40pmol, first probe 2.5pmol, second probe 5pmol, third probe 7.5pmol and fourth probe 8.75pmol; wherein, 1×PCR buffer includes: Tris-HClpH8 .5 67mM, (NH4) 2 SO 4 16 mM and 0.1% (w / v) Tween.

[0036] The first probe is shown in SEQ ID NO 01, its 5' ends are respectively labeled with a fluorescent group CY5, and its 3' ends are both marked with a quencher group BHQ2.

[0037] The second probe is shown in SEQ ID NO 02, its 5' ends are respectively labeled with a fluorescent group HEX, and its 3' ends are both marked with a quencher group BHQ1.

[0038]...

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PUM

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Abstract

The invention discloses an NAT2 (N-acetyltransferase-2) gene polymorphism fluorescence PCR (Polymerase Chain Reaction) melting curve detection kit, which comprises a first probe, a second probe, a third probe, a fourth probe, a positive primer and a reverse primer, wherein different fluorescence groups are respectively marked at 5' ends of the first probe, the second probe, the third probe and the fourth probe; and corresponding quenching groups are marked at the 3' ends of the first probe, the second probe, the third probe and the fourth probe. The detection of four mononucleotide polymorphisms on four sites can be completed in a single-tube PCR system; the sample genotype can be known through once fluorescence PCR melting curve analysis after the PCR amplification is completed; the whole operation can be completed in 3 hours; and the time consumption is low.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a NAT2 gene polymorphism fluorescent PCR melting curve detection kit. Background technique [0002] Tuberculosis is a disease caused by Mycobacterium tuberculosis infection, which is a serious problem plaguing human health all over the world. Mycobacterium tuberculosis can be transmitted through the air. At present, about 2 billion people in the world are infected with Mycobacterium tuberculosis. If not treated in time, each tuberculosis patient can infect 10 to 15 people per year on average. Tuberculosis is one of the infectious diseases that cause the largest number of deaths. There are about 8 to 10 million new tuberculosis patients worldwide each year, and about 2 to 3 million deaths due to tuberculosis each year, of which 95% are tuberculosis patients and 98% are tuberculosis patients. TB deaths occur in developing countries. Since isoniazid (INH) ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6876C12Q2600/156C12Q2527/107C12Q2563/107
Inventor 郭晓彤黄海荣宋娜杰
Owner XIAMEN ZEESAN BIOTECH
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