Amplification was performed in an automated thermal cycler programmed for one cycle of
94ºC for 5 minutes followed by 35 cycle reaction profile
involving 30 seconds of denaturation at 94ºC, 30 seconds
of annealing at 55ºC, and 1 minute and 30 seconds of
extension at 72ºC and single cycle of final extension at
72ºC for 7 minutes