Deep RNAseq of Viscum album
Viscum album leaves were flash frozen in liquid nitrogen and stored at -80C until processing. Total RNA was extracted from approximately 100 mg of leave and stem tissue of Viscum spp. using the Spectrum Total Plant RNA isolation kit (Sigma-Aldrich). For ibrary preparation and Next Generation Sequencingwe used the SENSE mRNA Library Preparation Kit for Illumina Sequencing (Lexogen, Vienna), following the manufacturers instructions. The protocol uses oligo-dT magnetic beads to target the poly-adenylated RNA molecules of the total extracted RNA, thus removing ribosomal and transfer RNA molecules, as well as messenger RNA molecules of prokaryotic origin. The purified mRNA was quantified in a Qubit 2.0 (Life Technologies) using the RNA assay kit. Illumina adapters were attached as the protocol instructed, after running a qPCR to determine the optimal number of amplification cycles.