Comparative evaluation of five different methods for DNA extraction from semen of buffalo bull

EVALUATION OF DNA EXTRACTION METHODS FROM SEMEN OF BUFFALO BULL

Authors

  • RAJNI KUMARI DIVISION OF LIVESTOCK AND FISHERY MANAGEMENT, ICAR-RCER, PATNA
  • KANISHT BATRA Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125004, India
  • VINAY KUMAR Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125004, India
  • AMAN KUMAR Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125004, India
  • SUSHILA MAAN
  • TRILOK NANDA Dept. of Animal Biotechnology, LUVAS, Hisar

Abstract

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           

Author Biographies

RAJNI KUMARI, DIVISION OF LIVESTOCK AND FISHERY MANAGEMENT, ICAR-RCER, PATNA

DIVISION OF LIVESTOCK AND FISHERY MANAGEMENT, ICAR-RCER, PATNA

KANISHT BATRA, Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125004, India

Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125004, India

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Published

2019-03-01

Issue

Vol. 6 No. 1 (2019): Journal of AgriSearch

Section

Articles

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