2.2. Preparation of DNA templates
Bacterial isolates were grown overnight on TSA at 37 C;
one isolated colony was picked and suspended in 100 ml sterile
distilled water and lysed by boiling for 15 min, followed by
freezing and subsequently centrifuged at 14,000 rpm for
15 min to pellet the cell debris. The supernatant was collected
and used as a template for amplification reaction. The DNA
concentration of the supernatants was measured using a
Nanodrop ND-1000 spectrophotometer (Thermoscientific,
Wilmington, DE) and adjusted to be approximately 100 ng/ml.