Veterinary Archives, Vol. 86 No. 6, 2016.
Original scientific paper
Molecular characterization of the mitochondrial 16S rRNA gene of cattle, buffalo and yak.
Deep P. Saikia
; Department of Animal Biotechnology, CVSc, AAU, Guwahati, Assam, India
Dhruba J. Kalita
; Department of Veterinary Biochemistry, CVSc, AAU, Guwahati, Assam, India
Probodh Borah
; Department of Animal Biotechnology, CVSc, AAU, Guwahati, Assam, India
Satya Sarma
; Department of Veterinary Biochemistry, CVSc, AAU, Guwahati, Assam, India
Rupam Dutta
; Department of Animal Biotechnology, CVSc, AAU, Guwahati, Assam, India
Deekshita Rajkhowa
; Department of Veterinary Biochemistry, CVSc, AAU, Guwahati, Assam, India
Abstract
A combination of polymerase chain reaction (PCR) with restriction fragment length polymorphism (RFLP) and nucleotide sequencing is the most preferred and efficient method for characterization of different species, in terms of detection power and applicability to large scale screening. The present study was carried out with the aim of developing the molecular fingerprint of the mitochondrial 16S rRNA gene of Cattle, Buffalo and Yak. Blood samples were collected randomly from ten different animals of each species for mitochondrial DNA extraction. The extracted DNA was used for the amplification of the 16S rRNA gene using universal primers. The size of the amplified products was 600bp. RFLP studies were carried out by digesting the amplicons using restriction enzymes viz. AluI, HinfI and HaeIII. The resulting RFLP pattern could easily identify and differentiate each of the species. Sequencing of the amplicons in all three species was carried out to confirm the variations at nucleotide level. Sequence analysis of the 16S rRNA gene using MEGA4 software and also PCRRFLP revealed that the 16S rRNA gene can be used as a good candidate for a molecular marker.
Keywords
PCR-RFLP; mitochondrial DNA; nucleotide sequence; 16S rRNA gene
Hrčak ID:
169828
URI
Publication date:
22.11.2016.
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