Comparative assessment of five distinct DNA extraction protocols from frozen buffalo semen

To enhance milk production in terms of quality and quantity, disease resistance, draught capacity, number of lactations, and so on, superior bulls and cows must be selected using genetic markers.Polymerase Chain Reaction (PCR) is the first step in the analysis of genetic markers and their incorporation into the population [13].PCR is a relatively quick, sensitive, and simple technique that allows for the quick amplification and in vitro examination of specific DNA sequences.The DNA template, quantity, and quality are necessary for the PCR process to be effective [14,21].Due to extensive artificial insemination programs, semen is a frequent source of the transfer of viral, bacterial, and parasite infections to cattle and buffaloes [7].Therefore, a feasible strategy for reducing the prevalence of these illnesses is to ABSTRACT: Selection utilizing genetic markers is a key aspect in achieving genetic improvement in populations.Genomic and proteomic analyses require a sufficient quantity of good-quality DNA, which can be isolated from animal sperm, hair, blood, tissues, saliva and cartilage.Due to the presence of solvents and dilutions in frozen semen, the physical and chemical properties of sperms and the non-cellular percentage of ejaculate, DNA extracted is often of poor quality.As per the literature, a wide variety of DNA extraction methods from semen was available.In the present study, five different extraction procedures were adopted to assess the quality and amount of DNA extracted from frozen semen straws.All the methods were found to obtain DNA, which was verified by nanodrop readings; however, the genomic DNA obtained by method I could not be visualized on 1% agarose gel.The DNA quality and quantity, obtained from these methods differ significantly (p<0.01).Successful amplification of the target sequence by PCR was observed with DNA extracted using the III and V methods.In conclusion, method V (modified high salt method) is the best procedure to obtain good quality and quantity DNA from buffalo frozen semen straws without any constraints in PCR.