Application of PCR-RFLP technique to determine Booroola gene polymorphism in the Sangsari sheep breed of Iran

Abstract

Phenotypic evaluation and the culling of candidate animals for traits by applying traditional animal breeding are usually costly tasks, which need to be carried out over a considerable span of time. The use of molecular genetics as an alternative method enables animal breeders to select eligible animals for the desirable trait, or traits, at earlier ages. Selection based upon markers could result in increased accuracy as well as a better selection response of animals. The Booroola gene has long been recognized as a candidate gene responsible for a higher rate of ewe ovulation, which results in increased litter size. In this study, blood samples were taken from 150 Sangsari sheep breed (140 ewes and 10 rams) in the Damghan animal breeding center using Venojects treated with an anticlot substance (EDTA). The sample DNA contents were salted out and extracted. After extraction, and after undertaking quantitative and qualitative tests (including spectrophotometry and gel agarose, 8%), the required amounts of DNA for polymerase chain reaction (PCR) were determined. Using the relevant primer, the related part was reproduced (190 bp) and then the PCR products were cut by AvaΠ enzymes for the gene, and 2 parts (30 bp and 160 bp) were produced for the target site. The results obtained in this study revealed that only the wild-type allele (+) was observed in the samples, indicating that, most likely, only genotype ++ was present in the population under consideration.