Development and optimization of cost-effective Tetra-primer ARMS PCR technique for detection of Myostatin gene g + 6723G>A polymorphism in Karakul sheep

Myostatin, also called growth and differentiation factor 8 (GDF8), acts as a negative regulator of muscle growth and development; Therefore, its role in animal growth and meat production is very important. Functional studies showed that a single nucleotide polymorphism in the 3'UTR region of this gene causes the elimination of its growth inhibitory role and finally the occurrence of double - muscled sheep. The tetra primer ARMS PCR method is an easy, fast, reliable and cost-effective method for the detection of nucleotide polymorphism. The present study was conducted with the aim of identifying single nucleotide polymorphism of myostatin gene (g+6723G>A) using tetra primer ARMS PCR technique in 86 Karakul sheep. Polymerase chain reaction was optimized using 3 control samples including 1 homozygous wild-type, 1 heterozygous and 1 homozygous mutant genotype of sharole breed sheep. All the studied Karakul sheep had the G allele, and the mutant type allele (A allele) was not observed, so the frequencies of the mutant type (A) and the wild type (G) alleles were 0 and 100%, respectively. To evaluate the accuracy of tetra primer ARMS PCR technique in this locus, all tested samples were also genotyped by PCR-RFLP method. The results indicated that two methods are the same as therefore using Tetra Primer ARMS PCR method been as economical to genotyping