RNA-Seq of Dibothriocephalus dendriticus plerocercoid
Adults of D. dendriticus were obtained from the body cavity of the Baikal omul, Coregonus migratorius. Total RNA from 0.5 to 1.0 g of tissue was isolated with TRIzol reagent (Ambion) and purified with simultaneous treatment with DNAase I on PureLink RNA Mini columns (Invitrogen). RNA quality was determined on a BA2100 Bioanalyzer using an RNA Nano Kit.The TruSeq Stranded mRNA Library Preparation Kit (Illumina) was used to prepare directional barcoded transcriptome libraries with double indexed UD according to the manufacturer's protocol, with modifications made to obtain longer embeddings (200-500 bp). 1 ?g of total RNA was collected, and the mRNA fragmentation time was 4 min. After amplification of the libraries, additional length selection was performed on AMPureXP magnetic particles - 0.65 volumes of AMPureXP were added to the diluted library. The quality and molarity of the obtained libraries were determined on a BA2100 bioanalyzer; the libraries were mixed in equimolar amounts to a total concentration of 2 nM. The obtained libraries were sequenced on an Illumina NextSeq550 High Throughput Sequencer using a NextSeq 550 High Output v2 Kit (300 cycles) in 150 bp paired-end reads.