total RNA was harvested using a Qiagen Rneasy mini kit (Cat# 74106) following the manufacturer protocol and including the on-column DNA digestion. 2 ug of total RNA was used as starting material for library construction using the KAPA Stranded mRNA Seq Kit (Roche Cat# 07962193001) following manufacturer protocol for a desired insert size of 100-200 bp. Briefly RNA was enriched for the Poly-A mRNA and reverse transcribed to double-stranded cDNA. The cDNA was end repaired, adenylated, and ligated with Illumina TruSeq adapters/indexes prior to 9 cycles of PCR. Final library quality was assessed on an Agilent TapeStation 2200. Libraries were quantified using KAPA Universal Library Quantification Kit (Roche Cat# 07960140001) before pooling libraries in equamolar concentrations. Sequencing was performed on Illumina HiSeq 4000 using single end, 50bp reads.
GEO Accession
GSM3887778