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Primers, probes and kit for detecting C-kit gene K642E mutation site

A K642E, mutation site technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of long C-kit gene mutation detection cycle, complicated result interpretation, and complicated operation process, etc. problem, to achieve the effect of safety, non-toxicity, low cost, short experimental period and simple operation

Inactive Publication Date: 2017-06-20
西安医臻生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The object of the present invention is to provide a primer, probe and kit for detecting the K642E mutation site of the C-kit gene, to solve the problem of the existing C-kit gene mutation detection cycle is long, the operation process is cumbersome, and the result interpretation is complicated. , low sensitivity, easy to pollute the problem

Method used

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  • Primers, probes and kit for detecting C-kit gene K642E mutation site
  • Primers, probes and kit for detecting C-kit gene K642E mutation site
  • Primers, probes and kit for detecting C-kit gene K642E mutation site

Examples

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Effect test

Embodiment 1

[0071] Example 1 A kit for detecting the K642E mutation site of the C-kit gene

[0072] The kit includes quantitative reaction system reagents and detection reaction system reagents, PCR buffer, dNTPs, MgCl 2 , TaqDNA polymerase and template DNA.

[0073] The quantitative reaction system reagents include upstream primers for specifically amplifying the target sequence, downstream primers for specifically amplifying the target sequence, probes for detecting the K642E mutation site of the C-kit gene, quality control Primer pairs and quality control probes; the detection reaction system reagents include peptide nucleic acid PNA probes for blocking wild-type gene amplification, upstream primers for specific amplification of target sequences, and specific amplification Downstream primers for targeting sequences, probes for detecting C-kit gene K642E mutation sites, quality control primer pairs and quality control probes.

[0074] The detection reaction system reagent comprises th...

Embodiment 2

[0084] The kit includes quantitative reaction system reagents and detection reaction system reagents, PCR buffer, dNTPs, MgCl 2 , TaqDNA polymerase and template DNA.

[0085] The quantitative reaction system reagents include upstream primers for specifically amplifying the target sequence, downstream primers for specifically amplifying the target sequence, probes for detecting the K642E mutation site of the C-kit gene, quality control Primer pairs and quality control probes; the detection reaction system reagents include peptide nucleic acid PNA probes for blocking wild-type gene amplification, upstream primers for specific amplification of target sequences, and specific amplification Downstream primers for targeting sequences, probes for detecting C-kit gene K642E mutation sites, quality control primer pairs and quality control probes.

[0086] The detection reaction system reagent comprises the sequence of the peptide nucleic acid PNA probe used to block wild-type gene ampl...

Embodiment 3

[0096] The kit includes quantitative reaction system reagents and detection reaction system reagents, PCR buffer, dNTPs, MgCl 2 , TaqDNA polymerase and template DNA.

[0097] The quantitative reaction system reagents include upstream primers for specifically amplifying the target sequence, downstream primers for specifically amplifying the target sequence, probes for detecting the K642E mutation site of the C-kit gene, quality control Primer pairs and quality control probes; the detection reaction system reagents include peptide nucleic acid PNA probes for blocking wild-type gene amplification, upstream primers for specific amplification of target sequences, and specific amplification Downstream primers for targeting sequences, probes for detecting C-kit gene K642E mutation sites, quality control primer pairs and quality control probes.

[0098] The detection reaction system reagent comprises the sequence of the peptide nucleic acid PNA probe used to block wild-type gene ampl...

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PUM

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Abstract

The invention discloses primers, probes and kit for detecting a C-kit gene K642E mutation site. The primers, probes and kit comprise a peptide nucleic acid (PNA) probe used for preventing amplification of wild type genes, an upstream primer used for specific amplification of targeting sequences, a downstream primer used for specific amplification of targeting sequences, a probe for detecting the C-kit gene K642E mutation site, a quality control prime pair and a quality control probe. The kit is suitable for various clinical sample types and has the remarkable advantages that specificity is high, sensitivity is high, the experimental period is short, operation is easy, safety is realized, toxicity is avoided, and the cost is low.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer, a probe and a kit for detecting the K642E mutation site of the C-kit gene. Background technique [0002] The C-kit proto-oncogene is located on the long arm of chromosome 4, with a total gene length of about 80 kb and composed of 21 exons. The mRNA of C-kit is 5185bp long, and its open reading frame is 2931bp long, encoding a transmembrane protein molecule of 976 amino acid residues, that is, the transmembrane tyrosine kinase receptor C-kit protein. The molecular weight of KIT protein is about 145kD. It is a member of the type III tyrosine kinase transmembrane receptor family. Region (signaling, amino acids 520-543) and a cytoplasmic region containing tyrosine kinase activity (amino acids 544-976). [0003] C-kit gene abnormalities are most common in acral and mucosal melanomas. These characteristics have laid a theoretical foundation for the individualized treatment of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/156
Inventor 李鹏王硕张晓飞张凤龙
Owner 西安医臻生物医药科技有限公司
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