tokens: Thermocycling conditions for both the unmethylated - and methylated - specific PCRs were an initial step of 5 minutes at 95 degrees C followed by 35 cycles of 30 sec . at 94 degrees C , 30 sec . at 60 . 5 degrees C and 60 sec . at 72 degrees C before a final elongation step at 72 degrees C for 7 min .