Primer test on real tissue
We first tested whether the designed primers (Table 1) could amplify DNA in simple PCRs conducted on 7 tissue samples of 6 different phyla: a nematode (Ascaridia galli ), cestode (Raillietina cesticillus ), insect (Apis mellifera ), arachnid (Ischyropsalis pyrenaea ), leech (Helobdella stagnalis ), gastropod (Cepaea hortensis ) and bird (Scolopax rusticola ). DNA extraction from tissues was performed using the kit DNeasy® Blood & Tissue Kit (Ref. 69506) (Quiagen). All PCR reactions (20 µl) were carried out using 1 µl of DNA in reactions with 1.25 µl of MgCl (25 mM), 2 µl of dNTPs (20 mM), 0.25 of 10X Buffer B, 0.3 µl of each forward (20 pM) and reverse (20 pM) primers, 1 µl of BSA (10mg/ml) and 0.25 µl of BIOTAQ™ DNA Polymerase of Bioline (ref. BIO-21060; 5 U/µl). The thermocycle conditions were: 96°C for 1 minute and 35 cycles of 94°C for 30 seconds, 57°C for 30 seconds, 72°C for 1 minute. PCR amplification capacity was confirmed by electrophoresis in 1.5% TBE agarose gel.