The PCR program for selective amplification was performed under the following conditions: 3 min at 94 C, followed by 12 cycles of 45 s at 94 C, 45 s at 65 C and 1 min at 72 C; the annealing temperature was decreased by 0.7 C per cycle from a starting point of 65 C during this stage (Touchdown), with a final round of 25 cycles of 94 C for 45 s, 56 C for 45 s, 72 C for 1 min and a final extension of 72 C for 1 min. MSAP and S-SAP molecular markers were used under the same conditions.