Comparative evaluation of five different methods for DNA extraction from semen of buffalo bull
EVALUATION OF DNA EXTRACTION METHODS FROM SEMEN OF BUFFALO BULL
Successful DNA extraction is indispensable for polymerase chain reaction (PCR) based molecular. Success of PCR method depends on the quality and quantity of the DNA template. There are many methods currently available for DNA purification from semen; but, problems related to contamination with foreign DNA, PCR inhibitors, and susceptibility to fragmentation are very common. In the present study, five different DNA extraction procedures were examined to ascertain their relative effectiveness for extracting DNA from semen samples of buffalo bull. The five methods included- two commercially supplied kits (Qiagen DNeasy Blood and Tissue Kit, Purelink Invitrogen), modified Qiagen DNeasy Blood and Tissue Kit method, Chelex 100 method and Phenol-chloroform with modified lysis buffer method. The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrophotometric measurements, real-time PCR amplifications and gel electrophoresis. Results revealed significant differences among the five procedures. Modified Qiagen DNeasy Blood and Tissue Kit method was found to be most appropriate for extracting high quality and suitable quantity of DNA from semen of buffalo bull.
Keywords: DNA extraction, semen, buffalo bull, evaluation