RNA Seq from pools of Ixodes ricinus adult male, female and nymph
Total RNA from single females, single males, and single nymphs was extracted from the Qiagen RNeasy Mini Kit, along with an incolumn DNase treatment. Two micrograms of the total RNA were taken and then reconstituted in the polyA binding buffer, and polyA messenger RNAs were purified from this pool by Oligo d(T)25 magnetic beads from NEB. A hundred nanogram aliquot of poly-A purified mRNA was used to prepare a long-read sequencing library using the Oxford Nanopore PCR-cDNA Sequencing Kit (SQK-PCB109; Oxford Nanopore Technologies, Oxford, UK).