Metatranscriptome of rotten Fomes fomentarius fruiting body
Prior to DNA/RNA extraction, the sample was homogenized in liquid nitrogen by mortar and pestle. Total RNA was extracted, in triplicate, from 200 mg of sample material using the NucleoSpin RNA Plant kit (Macherey-Nagel) according to manufacturers protocol after mixing with 900 l of the RA1 buffer and shaking on FastPrep-24 (MP Biomedicals) at 6.5 m.s1 twice for 20 s. Triplicates were pooled and treated with OneStep PCR Inhibitor Removal kit (Zymo Research). The DNA was removed using Turbo DNA-free kit (Invitrogen) and the efficiency of DNA removal was confirmed by PCR with the bacterial 515F (5-GTGCCAGCM GCCGCGGTAA-3) and 806R (5-GGACTACHVGGGTWTCTAAT-3) primers (Caporaso et al. 2012). Bacterial rRNA was depleted with the MICROBEexpress kit (Ambion). Eukaryotic, cytoplasmic and mitochondrial rRNA depletion as well as sequencing library preparation were done using the Trueseq Stranded Total RNA Gold kit (Illumina) following the manufacturers instructions. The RNA quality was, frequently, assessed using a 2100 Bioanalyzer (Agilent Technologies). The libraries were sequenced on an Illumina HiSeq 2500 (2  250 bases) at Brigham Young University Sequencing Centre, USA.