RNA library
The TruSeq Stranded Total RNA Kit (Illumina) was used for RNA library preparation. The rRNA depletion step was omitted. 80 ng of the total RNA (in 5µl volume) was combined with 13 µl of ‘Fragment, Prime and Finish mix’, for the RNA fragmentation step according the Illumina TruSeq stranded mRNA sample preparation guide. Subsequent steps were performed as described in the sample preparation guide. The library was sequenced on a MiSeq instrument; with v3 sequencing chemistry in 2x75 cycles paired end runs.