Raw reads (pair end) for transcriptome of vertebrate stages of Schistosoma japonicum
In this study, we analyzed the transcriptome of Schistosoma japonicum to discover alternative splicing events in this parasite, by applying RNA-seq to cDNA library of adults and schistosomula. Results were validated by RT-PCR and sequencing.Total RNA was isolated using TRIzol reagent (Thermo Scientific), following by RNA purification by RNeasy MiniElute Cleanup Kit (Qiagen), according to the manufacturer’s instructions. For library construction, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads (Illumina). The purified mRNA was cut into fragments with an average length of 155 bp using divalent cations at 95°C for 8 min. The cleaved mRNA fragments were reverse-transcribed into first strand of cDNA using random hexamers, prior to the synthesis of the second strand of cDNA. After end repair process, cDNA fragments were linked with adaptors and amplified by PCR. Equal amount of cDNA libraries of schistosomula and adults were mixed and the final concentration of this mixed library was adjusted to 2 nM. The sample was then sequenced on an Illumina HiSeq 2000 system.