Following an initial denaturation at 95°C for 10 min, 45 cycles each of 15 s denaturing at 95°C and 1 min annealing/elongation step at 60°C were used in qPCR.
The qRT-PCR reaction consisted of an initial denaturation at 94 °C and 40 amplifications cycles of 5 s at 94 °C and 60 s at 64 °C.
Cycling was started after an initial denaturation at 95°C for 5 min and ended with a final extension at 72°C for 5 min. Only the annealing temperature was changed for the experiments at different annealing temperatures.
After an initial denaturation at 95°C for 5minutes, PCR amplification was conducted for 20 35 cycles, and GAPDH was used as an internal control.
Reverse transcription was performed at 50°C for 30 min. After an initial denaturation at 94°C for 2 min, two amplification steps were performed.
PCR conditions included an initial denaturation at 95°C for 5 min followed by 20 amplification cycles (95°C for 30 s, 45°C for 30 s and 72°C for 1 min).
PCR involved an initial denaturation at 95°C for 15 min, 44 cycles of 10 sec at 94°C, 20 sec at 58°C, and 30 sec at 72°C.
DNA was amplified by an initial denaturation at 94°C for 5 min followed by 30 cycles of 30 seconds at 94°C, 1 min at 56°C and 30 seconds at 72°C.
Amplification was carried out using the 7300 Real-Time PCR Detection System (Applied Biosystems) with an initial denaturation at 95°C for two minutes followed by 50 cycles at 95°C for 30 seconds and 62°C for 30 seconds.
Conditions for PCR were an initial denaturation at 95°C for 3 min, 36 cycles of: 95°C 1 min, 50°C 1 min, 72°C 2 min, and a final extension for 7 min at 72°C.
PCR thermocycling conditions were an initial denaturation at 95°C for 7.5 minutes followed by repeated cycles (details below) of 95°C for 15 seconds, 52°C for 45 seconds and 72°C for 45 seconds.
