Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA extraction was performed using the QIAGEN RNeasy 96 kit with DNase digestion as per the manufacturer's instructions. All samples used for analysis had an RNA integrity number >8. The quantity and quality of total RNA were assessed by spectrophotometric analysis and by Agilent Tapestation 2200. Total RNA samples were normalized and randomized for further processing. Sequencing libraries were generated from 150ng total RNA using the Illumina TruSeq Stranded Total RNA sample preparation kit with Ribo Zero Gold as per manufacturer's instructions, quantified using the Kapa Library Quantification kit (Kapa Biosystems), normalized to 2nM and pooled as per the randomization plan. Pooled libraries were sequenced on a Illumina HiSeq 2500 sequencer for 2 × 50 cycles using the TruSeq PE Cluster Kit and TruSeq SBS Kit sequencing reagents (Illumina). Each lane was spiked with the PhiX Control library (Illumina) at a final concentration of 1 % (v/v) as a sequencing control