Polyadenylated RNA samples from adult male and female S. japonicum parasites were isolated from total RNA using oligo-(dT) conjugated magnetic beads (Dynabeads®, Invitrogen, CA, USA). The mRNA was interrupted into short fragments by adding the fragmentation buffer provided by the manufacturer (Illumina RNA-seq kit, part no. 1004898). With these short fragments as templates, random hexamer primers were used to synthesize the first-strand cDNA. The second-strand cDNA was synthesized using buffer, dNTPs, RNase H, and DNA polymerase I, respectively. Short fragments were purified following instructions accompanying the kit (QiaQuick PCR Purification Kit, Qiagen, Germany), and double-stranded cDNAs were end-repaired according to manufacturer-recommended protocols, followed by connection with Illumina adapters (Illumina RNA-seq kit, part no. 1004898). The fragments were first amplified by PCR. Purified cDNA fragments were pooled and indexed and loaded onto one lane of an Illumina GA IIX flow cell. A total of 75 pair-end sequencing cycles were carried out. Cluster formation, primer hybridization, and pair-end sequencing were performed according to the provided protocols
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