Quick detection method for Notch1 gene mutation

Abstract

The invention provides a quick detection method for Notch1 gene mutation, which comprises the following steps that a pair of wild type primers is designed at the two ends of a Notch1 gene mutational site; a pair of mutational type primers is designed at the mutational site; the wild type primers and the mutational type primers of Notch1 and Notch1-6 are designed respectively, the primers are amplified to be a wild type template and a mutational type template respectively; the two templates are subjected to PCR (polymerase chain reaction) amplification by the wild type primers to be a corresponding wild type target fragment and a mutational type target fragment respectively; the two fragments are mixed in different proportions; and an HRM (high resolution melting) method is adopted for distinguishing. Compared with other methods, the quick detection method has the advantages of high specificity, high sensitivity, convenience and quickness, and is higher in flux and lower in single cost.

Description

Technical field: [0001] The invention relates to the fields of biotechnology and medicine, especially molecular biology, molecular diagnosis and real-time quantitative PCR technology. Background technique: [0002] The Notch gene was first discovered in Drosophila, and its homologous gene in mammals encodes a single transmembrane receptor protein with a molecular weight of about 300KD; and the Notch information pathway includes DSL / Jagged ligand protein, and Components such as the cofactor CSL with DNA binding activity (4). When the ligand molecule combines with the Notch molecule on the adjacent cell membrane, Notch is degraded by protease, releasing the cytoplasmic region ICN (Intracellular domain of Notch) with nuclear localization signal, entering the nucleus and binding with CLS to regulate gene expression; this process It can be summarized as follows: ligand→Notch→enzyme cleavage→ICN→enter the nucleus→form CLS-ICN complex→promote gene transcription. The target genes ...

AI Technical Summary

Problems solved by technology

[0014] First: the sensitivity is not high, and a low proportion of mutations (<20%) cannot be detected;

[0015] Second: it takes a long time, usually 2-3 days;

[0016] Third: high cost

Images