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 a target gene, wherein the fluorescent signal probe is completely complementary to a mutant gene of the target gene and forms a single-base mismatch with a wild-type gene of the target gene; S2,adding a target gene sample to be detected and lambda exonuclease into a fluorescent signal probe-containing solution to form a reaction system; S3, acquiring a real-time fluorescence curve of the reaction system during the reaction, and judging whether the single-base mutation exists in the target gene sample to be detected. By the method, a single-base mutant gene sequence can be rapidly and accurately detected, and the selectivity to the single-base mutant gene sequence is high, so that the method can be applied to low-abundance mutation detection; in addition, by the method, the detectioncost is low, the preparation is easy, the operation is simple and the repeatability is high, and the method is easily popularized among non-professionals.

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...

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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

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