nematode 1
PCR amplification was performed in 25 L reaction mix containing: 1X Taq reaction Buffer (10 mM Tris-HCl at pH 8.3, 50 mM KCl and 1.5 mM MgCl2), 0.2 mM of each deoxynucleotide triphosphate, 0.5 pmol of each primer, 0.6 U of GoTaq DNA Polymerase and 10 ng of template DNA. PCR thermal profile was as following: an initial denaturation of 3 min at 95C followed by 35 cycles of 30 s denaturation at 95C, 30 s annealing at 52C and 1 min 20 s extension at 72C, with a final single extra extension step of 10 min at 72C. The obtained amplicon was Sanger sequenced and electropherograms assembled