PCR was carried out in a Biometra T3
thermocycler (Fisher Scientific) using the following cycling parameters:
ITS primers - pre-denaturation at 94 C for 5 min, 45
cycles of 94 C for 30 s, 57 C for 30 s, 72 C for 1 min and postelongation
of 72 C for 7 min. COI primers e pre-denaturation at
94 C for 5 min, 35 cycles of 94 C for 30 s, 50.9 C for 30 s, 72 C for
40 s and post-elongation of 72 C for 7 min. The products were
analyzed by ethidium bromide-stained 1.5% agarose gel electrophoresis.
PCR products were purified using the illustra™GFX™PCR
DNA and Gel Band Purification Kit (GE Healthcare) and sequenced
at Macrogen Inc. (South Korea) using the PCR primers.