Amplicon Illumina ITS1 rDNA and LSU rDNA sequences: Mixed Fasciola hepatica 90 eggs and Fasciola gigantica 10 eggs
DNA was amplified using ITS1 rDNA primers (S0922 F1 TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-AGTGCTAGGCTTAAAGAG; S0921 F2 TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-TTAAAGAGGAGATTTGGGC; S0923 R1 GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-CAAAGACCAGGTTATCAG) and LSU rDNA primers (S0925 F1 TCGTCGGCAGCGTCAGATGTGTATAA-GAGACAGGAGTAAACAGTGCGTGAAA; S0924 R1 GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-CTGAGAAAGTGCACTGACAAG) targeting specifically Fasciola spp. DNA. The ITS1 rDNA and LSU rDNA amplicons were used to evaluate SNPs within approximately 200 nt long amplicon. Sequencing using Illumina NGS platform was used to quantitatively evaluate the SNP using amplicons in the exponential phase of PCR. Pooled PCR amplicons were submitted for barcoding, library preparation Illumina Nextera XT V2 and sequencing on Illumina iSeq 100 System with 2x150 bp run at the Ramaciotti Centre for Genomics, University of New South Wales, Sydney, Australia.