A method for detecting single base mutation

Abstract

The invention relates to a method for detecting single base mutation in the field of gene mutation detection. The method comprises the following steps: S1, designing a fluorescent signal probe according to the target gene; the fluorescent signal probe is completely complementary to the mutant gene of the target gene, and forms a single base mismatch with the wild-type gene of the target gene; S2, Add the sample to be tested of the target gene and λ exonuclease to the solution containing the fluorescent signal probe to form a reaction system; S3, obtain the real-time fluorescence curve of the reaction system during the reaction, and judge whether the sample to be tested of the target gene is There is a single base mutation. The method of the invention can quickly and accurately detect the single-base mutation gene sequence, has high selectivity for the single-base mutation gene sequence, and can be used for low-abundance mutation detection. In addition, the detection cost of the method is low, easy to prepare, easy to operate, high in repeatability, and easy to be extended to non-professionals for operation.

Description

technical field [0001] The invention belongs to the technical field of gene mutation detection, and in particular relates to a method for detecting single base mutation. Background technique [0002] Gene sequencing technology has shown that base changes in the human genome are closely related to the formation and development of tumors. In a large number of isolated tumor patient tissue samples, single-base point mutations were found in the genomes of many tumor cells. However, in the early stage of tumor onset, the content of tumor cells is low, and the percentage of mutated genes in tumor cells is also relatively low. [0003] Gene mutation detection technology can be divided into sequencing type and non-sequencing type. Sequencing technology has high accuracy and can do repetitive sequences, but the cost of sample preparation is high, the concentration of nucleic acid required is high and the analysis time is long, and low-abundance detection cannot be performed, making...

AI Technical Summary

Problems solved by technology

The rate at which the lambda exonuclease degrades the double strand is sensitive to base pairing, and the presence of mismatches usually leads to a decrease in the degradation rate of the lambda exonuclease

Images