Primer, system and kit for MET gene amplification detection

A technology for detecting reagents and genes, which is applied in the field of primer sets, systems and kits for MET gene amplification detection, which can solve problems such as difficult detection of gene copy number, achieve short experimental operation cycle, reduce cross-contamination, and improve detection efficiency Effect

Active Publication Date: 2020-03-06
广州迈景基因医学科技有限公司
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Problems solved by technology

Moreover, the quantitative method in the present invention is not only applicable to the gene copy number detection of diploid animals, but also applicable to polyploid animals, plants, microorganisms, etc. to determine the copy number of a certain gene, thus solving the problem that the PCR method is not easy to detect. Th...

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  • Primer, system and kit for MET gene amplification detection
  • Primer, system and kit for MET gene amplification detection
  • Primer, system and kit for MET gene amplification detection

Examples

Experimental program
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Embodiment Construction

[0046] This kit needs to design the primers of the MET gene and the internal reference gene, and use the relative quantitative 2-△△Ct method to construct three plasmids, one as a calibration plasmid, and the other two as standard plasmids. The Ct values ​​of the MET gene and the internal reference gene were substituted into the formula to calculate the copy number of the MET gene.

[0047] Since the human genome is diploid, the copy number of the MET gene is 2 in the genome of normal cells. If the detection result is greater than 2, it indicates that there is amplification of the MET gene, and the specific copy number can be calculated. This is used to detect whether the MET gene is amplified.

[0048] 1. Primer Design

[0049] In this experiment, multiple pairs of primers were designed for the MET gene and the internal reference gene, and the optimal primer combination was screened out. Select an internal reference gene as a reference for the normal amplification of the sys...

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Abstract

The invention discloses a primer group for MET gene amplification detection, a kit and a related medication guidance system, and is characterized by constructing calibration plasmids and standard plasmids for correcting errors of primers and an amplification system, and the accuracy of the kit is improved. A 2-Ct method is adopted, is suitable for gene copy number detection of diploid animals, andis also suitable for determining the copy number of a certain gene of polyploid animals, plants, microorganisms and the like, so that the problem that PCR methods are not easy to detect the gene copynumber is solved. The method is simple and easy to operate, a result is easy to interpret, and a very beneficial thought is provided for other application of gene copy number detection (such as geneamplification and chromosome structural variation).

Description

technical field [0001] The invention relates to the field of biological technology, and relates to a primer set, system and kit for MET gene amplification detection. Background technique [0002] The morbidity and mortality of lung cancer rank first among malignant tumors in the world and in my country, and it is the main cause of cancer-related death. About 85% to 90% of lung cancers are non-small cell lung cancer (NSCLC). For patients with non-small cell lung cancer (NSCLC) with EGFR mutations, epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) Gefitinib and erlotinib are effective drugs for this type of lung cancer. [0003] Despite the enormous benefit of EGFR TKIs in EGFR-mutated non-small cell lung cancer (NSCLC), patients eventually developed resistance to both gefitinib and erlotinib (known as "acquired resistance"), studies have shown. Medicinal properties"). [0004] Two mechanisms of acquired resistance have been identified. The first me...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6886C12Q2600/106C12Q2600/118C12Q2600/158C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 闫文瑞林威唐同民蔡兴盛李梦真邓泱泱杨冬成
Owner 广州迈景基因医学科技有限公司
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