The mixture was heated in Programme Thermal Controller (Applied biosystem; PTC- 100™) using an initial hot start of 95°C for 15 minutes, followed by 35 cycles of 95°C for 1 minute, 55°C for 1 minute, and 72°C for 1 minute; a final extension step of 72°C for 10 minutes to complete the cycle.
The mixture was heated in a Thermal Cycler (Applied Biosystems; PTC- 100™) using an initial hot start of 95°C for 15 minutes, followed by 35 cycles of 95°C for 1 minute, 55°C for 2 minute, and 72°C for 1 minute; a final extension step of 72°C for 10 minutes to complete the cycle.
Cycle conditions were 95°C for 5 minutes, 35 cycles of 95°C for 1 minute, 60°C for 1 minute, 72°C for 1 minute and a final extension at 72°C for 10 minutes.
Standard PCR conditions were an initial cycle of 95°C for 2 minutes followed by 35 cycles of 95°C for 1 minute, 52°C for 1 minute, 72°C for 1 minute and a final cycle of 72°C for 10 minutes.
The reaction included an initial 5-min denaturation at 94°C; followed by 27 cycles of 94°C for 45 minutes, 60°C for 45 minutes, and 72°C for 1 minute; and a final 10 - minute extension at 72°C.
Amplification was performed using the following conditions: 94°C for 2 minutes, 30 cycles at 94°C for 1 minute, 60°C for 1 minute and 72°C for 1 minute, and a final cycle of elongation at 72°C for 4 minutes.
PCR conditions consisted of an initial denaturation step of 95°C for 5 minutes, followed by 24 cycles of 95°C for 1 minute, 58°C for 1 minute and 72°C for 1 minute, with a final extension of 72°C for 10 minutes.
Conditions followed were initial denaturation at 94°C for 4 minutes, followed by 35 cycles at 90°C for 1 minute, 60°C for 1 minute, and 72°C for 1 minute, and a final extension at 72°C for 10 minutes.
PCR was conducted using the following program: 94°C for 10 minutes, followed by 30 cycles of 94°C for 1 minute, 55°C for 1 minute and 72°C for 1 minute, and a final elongation step at 72°C for 10 minutes.
A denaturing step of 2 minutes at 95°C was followed by 35 cycles of 95°C for 30 seconds, 56°C for 30 seconds, and 72°C for one minute and a final 5 minute elongation step at 72°C.
PCR conditions for all primers sets were: denaturation at 94°C for 2 minutes, 30 cycles of denaturation at 94°C for 30 seconds and annealing and elongation at 68°C for 1 minute, and a final 10 minutes elongation at 72°C.
