JAK2 gene specific mutation detection kit and JAK2 gene specific mutation detection method based on double-stranded DNA peptide ligase

Abstract

The invention provides a JAK2 gene g.[146452_146455 delta ATGA] mutation detection kit and a JAK2 gene g.[146452_146455 delta ATGA] mutation detection method based on double-stranded DNA peptide ligase, wherein a target gene mutation fragment is directly captured and enriched before exponential amplification, and then the target gene mutation fragment is specifically amplified and detected. The JAK2 gene g.[146452_146455delta ATGA] detection kit mainly comprises dDPlaseII ligase, a dDPlaseII ligase buffer solution, RNase H enzyme, magnetic polypeptide Ps, an RNA primer Pr, a DNA primer Pn, a DNA fragment Pd and a DNA primer Pm, wherein the amino acid sequence of dDPlaseII ligase is represented by SEQ ID NO:1. The detection method mainly comprises the following four parts of pre-amplification, RNase H enzyme digestion, dDPlaseII enzyme linking and magnetic bead separation, and specific amplification. The detection kit and the detection method provided by the invention have the characteristics of strong specificity, accuracy, high efficiency, simplicity, convenience and economy, can effectively detect JAK2 gene g.[146452_146455delta ATGA] mutation with the content as low as 1% in a sample, and can be applied to prediction and diagnosis and treatment of pulmonary metastasis of male colorectal cancer patients.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a double-stranded DNA peptide ligase-based JAK2 gene g.[146452_146455delATGA] mutation detection kit and a detection method thereof. Background technique [0002] Gene is the genetic material of an organism, and the protein expressed by it bears various life functions in the organism. Mutations or mutations in genes will lead to different biological characteristics or disease states in organisms. Gene mutation is a change in the nucleotide composition or sequence of a gene in its molecular structure, mainly including point mutations, frameshift mutations, deletion mutations and insertion mutations. Gene mutations are closely related to the occurrence and development of tumors. At present, cancer has become the disease with the highest morbidity and mortality in the world. According to the data released by the National Cancer Center in 2019, there were about 3.929 million new c...

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Problems solved by technology

Most of them are PCR-based methods, however, conventional PCR amplification cannot distinguish wild-type genes from mutant genes, and requires specific design or analysis

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