Optimization of polymerase chain reaction method for identification of DGAT1 gene polymorphisms in Holstein cattle

The article presents the results of a molecular genetic study of the DGAT1 gene associated with fatty milk in cattle, performed using the polymerase chain reaction (PCR) method. The research was aimed at improving molecular genetic approaches to the analysis of the DGAT1 gene and determining the genetic structure of the Holstein cattle population by alleles A (alanine) and K (lysine). The study was conducted on the basis of the Krasnodarskoye experimental farm of the Kuban State Agrarian University, where blood samples were collected and further molecular genetic analysis was carried out. In the course of the study, DNA was successfully isolated from whole blood on the livestock of Holstein heifers of the Krasnodarskoye educational and experimental farm of the Kuban State Agrarian University, the quality of the isolated DNA met the established requirements for the ratio of optical densities A260/A280 and A260/A230, which confirms its suitability for research. Polymerase chain reaction conditions were optimized in order to increase the yield of amplicons. Allele-specific PCR made it possible to successfully amplify fragments of the DGAT1 gene with sizes of 368 and 244 bp, corresponding to the K (lysine) and A (alanine) alleles, respectively. The obtained results create prerequisites for the subsequent analysis of the distribution of the A and K alleles of the DGAT1 gene in the studied population and their possible use in breeding work aimed at increasing the dairy productivity of cattle.