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KRAS gene mutation detection method and kit

A detection kit and detection method technology, applied in the fields of genetic engineering and molecular biology, can solve the problems of low detection efficiency and complicated steps, and achieve the effect of simplifying experimental steps and improving experimental efficiency.

Active Publication Date: 2015-03-25
SHENGZHEN CHINA GENE TECH COMPANY
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Problems solved by technology

[0006] The purpose of the present invention is to provide a KRAS gene mutation detection method and kit, aiming to solve the problems of complicated steps and low detection efficiency in the prior art

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  • KRAS gene mutation detection method and kit

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[0036] The present invention proposes the first embodiment, and the present invention also provides a KRAS gene mutation detection kit, including KRAS-specific primers, and the KRAS-specific primers are used to specifically amplify the target region in the sample to be sequenced, and At least one of the KRAS-specific primers used to amplify each target region contains a cleavable site or excisable sequence.

[0037] It should be noted that after the KRAS-specific primers in the kit of the present invention amplify the target region in the KRAS gene, a breakable site or excisable sequence is brought into the amplified product, so that the amplified product is Subsequent cleavage and ligation can be performed in the same reaction system, which simplifies the steps of KRAS gene mutation detection and improves detection efficiency.

[0038] Wherein, the kit further includes a cleaving agent, which is used to specifically cleave the cleavable site or excisable sequence in the ampli...

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Abstract

The invention relates to the field of gene engineering and molecular biology, and provides a KRAS gene mutation detection method and a kit. The method comprises the following steps: A, amplifying a target region in a to-be-sequenced sample by using a KRAS specific primer to obtain an amplification product, wherein at least one PCR (Polymerase Chain Reaction) primer in the KRAS specific primer for amplifying each target region comprises a breakable site or a cuttable sequence; B, adding the amplification product into a cutting-connection reaction system to perform cutting and connection so as to connect a joint I and a joint II to the two ends of the amplification products to obtain library molecules; C, performing monomolecular amplification on the library molecules comprising the joint I and the joint II to obtain a monomolecular amplification product; D, performing high-throughput sequencing on the monomolecular amplification product to obtain sequence information on the target region. According to the method and the kit, the experiment steps can be simplified and the detection efficiency is improved.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, and more specifically relates to a KRAS gene mutation detection method and a kit. Background technique [0002] The ras gene family has three genes related to human tumors—HRAS, KRAS and NRAS, which are located on chromosome 11, 12 and 1, respectively. KRAS is also known as the p21 gene because it encodes a 21kD ras protein. Among the ras genes, KRAS has the greatest impact on human cancer. It is a molecular switch: when normal, it controls the pathways that regulate cell growth; when abnormal, it causes cells to continue to grow and prevents cells from self-destruction. It participates in intracellular signal transmission. When the KRAS gene is mutated, the gene will be permanently activated, unable to produce normal ras protein, causing intracellular signal transduction disorder, cell proliferation out of control and cancerous. Detection of KRAS gene mutation is an im...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q1/6886C12Q2600/156C12Q2535/122C12Q2525/131C12Q2521/301C12Q2525/191
Inventor 盛司潼钟茂春
Owner SHENGZHEN CHINA GENE TECH COMPANY
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