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Rs12979860 genotyping dual-color fluorescent PCR rapid detection kit

A detection kit and two-color fluorescence technology, applied in the biological field, can solve the problems of difficult deployment in general hospitals, expensive equipment, complicated operation, etc., and achieve the effects of high sensitivity, short detection time, and easy operation

Inactive Publication Date: 2017-02-15
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are many methods for identifying the polymorphism of the rs12979860 locus gene, mainly using methods such as gene sequencing, gene chip, high-resolution melting curve (HRM) and enzyme digestion, but most of the operations are complicated, time-consuming, and require The equipment is expensive and the specificity is poor, so it is difficult to carry out in ordinary hospitals

Method used

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  • Rs12979860 genotyping dual-color fluorescent PCR rapid detection kit
  • Rs12979860 genotyping dual-color fluorescent PCR rapid detection kit
  • Rs12979860 genotyping dual-color fluorescent PCR rapid detection kit

Examples

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Embodiment 1

[0024] This embodiment provides a rapid detection kit for rs12979860 locus genotyping two-color fluorescent PCR

[0025] 1. Kit composition and preparation

[0026] 1) Primers and probe sets

[0027] Primer probe set based on rs12979860 site

[0028] Forward primer F, the nucleotide sequence of which is shown in SEQ ID NO.1;

[0029] 5'CGGTCGTGCCTGTCGTGT3';

[0030] Reverse primer R, the nucleotide sequence of which is shown in SEQ ID NO.2;

[0031] 5'AGCGCGGAGTGCAATTCA 3';

[0032] C fluorescent probe S1, the nucleotide sequence of which is shown in SEQ ID NO.3;

[0033] 5'FAM-ACCCTGGTTCGCGCC-NFQ 3'-MGB, FAM probe;

[0034] T fluorescent probe S2, the nucleotide sequence of which is shown in SEQ ID NO.4;

[0035] 5'VIC-TGGTTCACGCCTTC-NFQ 3'-MGB, VIC probe;

[0036] The above primers and probes were all synthesized artificially by Shanghai Sangon Biotechnology Co., Ltd.

[0037] 2) DNA extraction solution

[0038] The DNA extraction solution can use a commercially av...

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Abstract

The invention discloses an rs12979860 locus genotyping dual-color fluorescent PCR rapid detection kit. According to the kit, a primer and a TaqMan-MGB probe are redesigned, a reaction system is optimized, bi-component hot-start DNA polymerase forms an enzyme activity automatic regulation system, ROX Reference Dye can eliminate a signal background and correct fluorescence signal errors between holes, therefore, the kit achieves accuracy, high amplification efficiency, high sensitivity, good specificity, good repeatability, easy and convenient operation and shorter detection time, and the technology can be applied and popularized clinically.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid detection kit for two-color fluorescence PCR based on rs12979860 site genotyping. Background technique [0002] Hepatitis C is a viral hepatitis caused by hepatitis C virus (HCV) infection, which is mainly transmitted through blood transfusion, acupuncture and drug abuse. Existing studies have shown that the rs12979860 polymorphism of the human IL-28B gene is correlated with the spontaneous clearance of HCV and the efficacy of antiviral therapy. [0003] At present, there are many methods for identifying the polymorphism of the rs12979860 locus gene, mainly using methods such as gene sequencing, gene chip, high-resolution melting curve (HRM) and enzyme digestion, but most of the operations are complicated, time-consuming, and require The equipment is expensive and the specificity is poor, so it is difficult to carry out in ordinary hospitals. Contents of the invention [...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2531/113C12Q2561/101
Inventor 赵友云孙莉军郑毅王业富
Owner WUHAN UNIV
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