PCR primer and method for detecting genotyping of SNP sites of ACVR1 gene

Abstract

The invention relates to the technical field of molecular detection and discloses a PCR primer and a method for detecting genotyping of SNP sites of an ACVR1 gene. The method comprises the following steps: design and synthesis of the PCR primer: designing the primer for PCR in NCBI according to a swine ACVR1 gene sequence predicted on NCBI, wherein the final concentration of the primer after dilution is 10 pmol / L; step 2, PCR amplification: taking muscular tissue DNA extracted in the step 1 as a template, performing PCR amplification by the primer according to the reaction procedure of PCR amplification; and step 3, reaction system amplification. According to the PCR primer and a method for detecting genotyping of SNP sites of the ACVR1 gene, internal contrast enzyme digestion sites are introduced into a PCR product, the defects that the used fast enzyme NdeI digestion efficiency of the PCR product is lower and incomplete enzyme digestion can be caused easily are overcome, the condition of incomplete enzyme digestion can be recognized according to an electrophoretogram, besides, the genotyping can be distinguished according to the condition of characteristic strip of each genotypeunder the condition of incomplete enzyme digestion, and accuracy of genotype distinguishing is guaranteed.

Description

technical field [0001] The invention relates to the technical field of molecular detection, in particular to a PCR primer and a method for detecting the SNP site genotyping of ACVR1 gene. Background technique [0002] The ACVR1 gene is essential for embryonic development and growth, involves multiple cells and tissue types, such as muscle, bone, and nervous system, and has temporal and spatial expression specificity during mouse development. Studies have shown that knocking out the ACVR1 gene causes mouse embryos to block early in development, so conditional mutants are often used to study the function of the ACVR1 gene in tissue / organ development. The current research on the ACVR1 gene is mainly focused on the research of progressive muscle ossification disease (Fibrodysplasia Ossificans Progressiva, FOP) and glioma (Diffuse Intrinsic Pontine Glioma, DIPG), and revealed the key mutations related to the two . [0003] At present, methods for detecting the genotype of ACVR1...

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies of the prior art, the present invention provides a PCR primer and method for detecting the SNP site genotyping of the ACVR1 gene. By introducing an internal control enzyme cutting site into the PCR product, the fast enzyme NdeI used to cut the PCR product is overcome. The efficiency of enzyme digestion is relatively low, and it is easy to cause the disadvantage of incomplete digestion. The situation of incomplete digestion can be identified according to the electrophoretic pattern of the digestion product, and in the case of incomplete digestion, it can still be detected according to the characteristic conditions of each genotype. The genotype can be identified according to the condition of the band, which ensures the accuracy of the genotype, and solves the problem that the current genotyping method of the SNP site of the ACVR1 gene requires high-quality personnel, cumbersome operations, and high testing costs.

Images