JAK1 gene specific mutation detection kit and detection method thereof

Abstract

The invention provides a JAK1 gene g.[227149insA] mutation detection kit based on double-stranded DNA peptide ligase and a method thereof. The invention is characterized in that a target gene mutationfragment is directly captured and enriched before exponential amplification, and then the target gene mutation fragment is specifically amplified and detected. The JAK1 gene g.[227149insA] detectionkit mainly comprises dDPlaseI ligase, a dDPlaseI 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 theamino acid sequence of dDPlaseI ligase is as shown in SEQ ID NO: 1. The detection method mainly comprises the following four parts: pre-amplification, RNase H enzyme digestion, dDPlaseI enzyme connection 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 JAK1 gene g.[ 227149insA] mutation with the content as low as 1% in a sample, and can be applied to prediction and diagnosis and treatment of liver metastasis of middle-aged female 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 JAK1 gene g.[227149insA] mutation detection kit and a detection method. 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 cancer cases in my ...

AI Technical Summary

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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