tokens: The PCR were performed with primers 1a and 1b using Advantage (R) 2 Polymerase Mix ( Clontech ) as follows : 95 degrees C , 1 min ; 5 cycles at 95 degrees C for 30 s and 68 degrees C for 1 min ; 30 cycles at 95 degrees C for 30 s , 58 degrees C for 30 s and 68 degrees C for 30 s , and finally 68 degrees C for 1 min .