Detection method of three-way crossbred wagyu cattle CAPN1 gene A316G mutation site and application thereof

Abstract

The invention provides a detection method of a three-way crossbred wagyu cattle CAPN1 gene A316G mutation site and an application. According to the detection method, with to-be-detected cattle whole genome DNA as a template, a cattle CAPN1 gene segment undergoes PCR amplification through a T-ARMS-PCR primer composition; and the genotype of the CAPN1 gene single nucleotide polymorphism site is identified in accordance with an agarose gel electrophoresis result. The T-ARMS-PCR primer composition comprises specific outer primers composed of sequences shown as SEQ ID No.1 and SEQ ID No.2 as well as site-specific primers composed of sequences shown as SEQ ID No.2 and SEQ ID No.3. The tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) detection method for three-way crossbredwagyu cattle CAPN1 gene A316G mutation, by conducting PCR amplification and agarose gel electrophoresis identification on the specifically designed primers, can be used for detecting single nucleotidepolymorphism of the site simply, rapidly and accurately with low cost; therefore the detection method is conducive to early screening of high-quality crossbred cattle and is capable of taking an assisting effect in improving the selection accuracy of three-way crossbred wagyu cattle of which meat quality traits are excellent.

Description

technical field [0001] The invention belongs to the field of animal breeding molecular biology detection, relates to the detection of gene single nucleotide polymorphism (SNP), in particular to a single-nucleus for detecting the C / G transversion of Ca-activated neutral protease 1 gene in Wagyu three-way hybrid cattle Nucleic acid polymorphism (SNP) four-primer amplification hindered mutation system PCR (T-ARMS-PCR) method. Background technique [0002] Existing studies have shown that Japanese Black Wagyu was used as the terminal male parent, Yili offal cattle (crossbreeding Limousin and Luxi cattle), west offal cattle (crossbreeding Simmental and Luxi cattle), summer offspring cattle (crossbreeding from Charolais) The wagyu three-way hybrid cattle crossed with Luxi cattle) as the female parent not only retains the advantages of easy breeding and strong adaptability of the female cow group, but also inherits the characteristics of the male parent Wagyu beef with obvious marb...

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Problems solved by technology

[0008] However, there is currently no report on the detection of the A316G mutation of the CAPN1 gene in Wagyu three-way hybrid cattle.

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