“…The polymerase chain reaction (
PCR) to confirm the identity of the nematode specie was carried out by the amplification of the internal transcribed spacer (
ITS) region, the D2D3 expansion segment of the 28S gene of the ribosomal DNA, and the portion of the cytochrome oxidase (
coxI ) gene of the mitochondrial DNA.
PCR of the
ITS region was carried out as described by
Chen et al 2005 using KAPA2G 40 Robust HotStart ReadyMix (KAPA Biosystems) with the primer combination of S-ITS1 (5'-TTGATTACGTCCCTGCCCTTT-3') and 28S (5'-TTTCACTCGCCGTTACTAAGG-3'). Amplification was carried out in a thermal cycler with the following cycling condition; 1 cycle at 94 °C for 4 min, followed by 30 cycles at 94 °C for 30 sec, 52 °C for 30 sec, and 72 °C for 2 min 30 sec, and ending with one cycle at 72 °C for 7 min and finally kept at 4 °C.…”