The Use of High Resolution Melting Analysis for Detection of the CAPN530 Polymorphism in Slovak Simmental Bulls

Abstract

The high-resolution melting analysis is a cost-effective and simple post-PCR technique that can be used for SNP genotyping. The objective of this study was to apply the high resolution melting analysis for detection of SNP polymorphism CAPN530 of bovine calpain gene (CAPN1) in Simmental bulls. A total 42 Simmental bulls were included in this study. Genomic DNA was extracted from sperm by commercial Nucleospin Tissue kit. The amplification and high resolution melting analysis (HRMA) was done by commercial SensiMixTM HRM kit in the real-time thermocycler Rotor-Gene 6000. The confirmation of sensitivity of this method was done by PCR-RFLP method. In the analyzed population of Simmental bulls were detected the following frequencies of alleles and genotypes for the SNP CAPN530 of the CAPN1 gene. Frequencies of allele A and allele G were 0.3929 and 0.6071 and frequencies of genotypes were 0.1191 (genotype AA), 0.5476 (genotype AG) and 0.3333 (genotype GG).

Keywords

• cattle
• genomic dna
• high-resolution melting analysis (hrma)
• snp polymorphism capn530