Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Sequencing libraries were created using Illumina’s TruSeqTM RNA sample preparation kit (San Diego, CA) according to manufacturer’s protocol. Total RNA was purified using oligo dT magnetic beads, fragmented, and reverse-transcribed using SuperScript II (Invitrogen, Inc., Carlsbad, CA) to synthesize first strand cDNA. After second strand synthesis, Illumina specific adapters containing unique barcodes were ligated to the ends of the double-stranded cDNA. Fragments containing adapters on both ends were then enriched and amplified with PCR, quantified with qPCR, and run on the Agilent Bioanalyzer DNA-1000 chip to estimate fragment size. Samples were then multiplexed and sequenced on the Illumina 2500. RNA libraries were prepared for sequencing using standard Illumina protocols