BUFFALO (Bubalus bubalis) SPECIFIC PRIMERS FOR CYTOCHROME B GENE FOR BUFFALO DNA DETECTION USING PCR METHOD

Abstract

This invention relates to the design of specific primers for buffalo (Bubalus bubalis) cytochrome b (Cytb) gene for the detection of buffalo DNA using conventional PCR method. The primer sequence of buffalo species specific Cytb gene consists of forward sequence 5'- TTAGTACTATTCGCACCCGACCTC-3' and reverse 5'- TCGTTGTTTGGATGTATGTAGCAG-3'. This primer sequence has an amplicon size of 216 bp. The primers were used in a conventional PCR process operated under the following conditions, initiation stage of denaturation at 95ºC for 60 seconds, denaturation stage at 95ºC for 15 seconds, annealing stage at 63ºC for 15 seconds, elongation at 72ºC for 10 seconds cycle, final elongation at 72ºC for 60 seconds and at 35 cycles. This invention aims to amplify buffalo species specifically using conventional PCR methods so that it can detect the presence of buffalo DNA in processed products, especially for processed buffalo skin crackers.