RNA-Seq of Schistosoma mekongi: adult male rep.2
Library was constructed using NEBNext  UltraTM RNA Library Prep Kit for Illumina  according to the manufacturer s protocol. Briefly, poly(A) mRNA isolation was performed using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB). The mRNA fragmentation and priming was performed using NEBNext First Strand Synthesis Reaction Buffer and NEBNext Random Primers. First strand cDNA was synthesized using ProtoScript II Reverse Transcriptase and the second- strand cDNA was synthesized using Second Strand Synthesis Enzyme Mix. The purified double-stranded cDNA was then treated with End Prep Enzyme Mix to repair both ends and add a dA-tailing, followed by a T-A ligation to add adaptors to both ends. Size selection of Adaptor-ligated DNA was then performed using AxyPrep Mag PCR Clean-up (Axygen), and fragments of ~360 bp were recovered. Sample was then amplified by PCR. The PCR products were cleaned up using AxyPrep Mag PCR Clean-up (Axygen), validated using an Agilent 2100 Bioanalyzer and quantified by Qubit 2.0 Fluorometer. Then library loaded on an Illumina HiSeq X-Ten instrument. Sequencing was carried out using a 2x150bp paired-end configuration.