title: Day 5 biological replicate 2
design: Large molecular weight RNAs were differentially isolated from the small molecular weight RNA fraction using the SPLIT  RNA extraction kit (Lexogen, Vienna, Austria) as per the manufacturer s instructions. RNA quality was evaluated using the Agilent RNA 6000 Nano Kit on a Bioanalyser 2100 (Agilent Technologies, Santa Clara, CA, USA). Only RNA samples with an integrity number > 7.0 were kept for RNA-seq library construction. Each sample was quantified using a Nanodrop 1000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA) and, prior to constructing RNA-seq libraries, 1.5  g of RNA was enriched in poly-A RNA using magnetic beads with poly-T oligonucleotides. Enriched poly-A RNA was used to construct libraries using the Illumina TrueSeq RNA sample prep kit v2 (llumina, San Diego, Ca, USA) as per the manufacturer s instructions except that the RNA fragmentation step was performed during six minutes.