title: RNA-Seq of Bombus terrestris: library Bter4
design: Head and abdomen dissection > totan RNA extraction (QIAgen RNeasy lipid tissue) + on-column DNase I > total RNA pooled per 5 individuals (10 µg each) > NanoDrop 2000c, Agilent TApeStation > Illumina TruSeq Stranded mRNA Library Preparation Kit (dilution to 10 ng/µl + 2rounds of polyA-mRNA selection using poly-T oligo attached magnetic beads on 500 ng total RNA + RNA fragmentation to 200-250 bp + cDNA synthesis + blunt ending + adapter ligation + PCR > Agilent TapeSTation D1000 > single lane HiSeq 2500 + rapid run mode + 2x100 bp read format (average 20M reads per sample).