The amplification reaction mixture
consisted of 1 PCR reaction buffer, 2.5 mM MgCl2, 0.12 mM
deoxynucleoside triphosphate, 0.2 mM of each primer, 1 U Taq DNA
polymerase and 1 ml of total DNA. The PCR program consisted of an
initial step at 94 C for 3 min, 35 cycles of denaturation at 94 C for
45 s, annealing for 1 min at 55 C and elongation for 2 min s
at 72 C, followed by final elongation step at 72 C for 8 min. The
ITS-PCR amplification patterns and 16S products were
migrated respectively on standard 2% agarose gels in
0.5 TriseborateeEDTA buffer and stained for 30 min in 0.5 mg/L
ethidium bromide solution. The amplified 16S rRNA fragments
were sequenced and identified by comparison with those available
at the National Centre for Biotechnology Information (NCBI) database