title: Fasciolopsis buski Transcriptome or Gene expression
design: the adult flukes were digested overnight in extraction buffer [containing 1% sodium dodecyl sulfate (SDS), 25 mg Proteinase K] at 37C, prior to DNA recovery. Genomic DNA was then extracted from lysed individual worms by ethanol precipitation technique or by FTA cards using Whatmans FTA Purification Reagent [33]. Quality and quantity of the DNA was assessed by 0.8% agarose gel electrophoresis (Sigma-Aldrich, USA), spectrophotometer (Nanodrop 1000, Thermofischer USA) and flurometer (Qubit, Thermofischer, USA). Whole genome shot-gun libraries for Illumina sequencing were generated and the genomic DNA was sheared (Covaris, USA), end-repaired, adenylated phosphorylate ligated to illumina adapters (TruSeq DNA, Ilumina, USA), to generate short (300-350 bp) fragment and long (500  550) bp insert libraries. Both libraries were amplified using 10 cycles of PCR KapaHiFiHotstart PCR ready mix (Kapa Biosystems Inc., USA) and adapter removal by Solid Phase Reversible Immobilization (SPRI) beads (Ampure XP beads, Beckmann-coulter, USA). Prior to sequencing, fragment analysis was performed (High Sensitivity Bioanalyzer Chip, Agilent, USA), quantified by qPCR. 36cycle sequencing was performed on Illumina Genome analyzer II (SBS kit v5, Illumina, USA) and Illumina HiSeq1000 (Illumina, USA) to generate 16 million 100 paired-end reads from the shot-gun library