Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was extracted using protocols modified from the Qiagen RNAprotect Bacteria Reagent Handbook. The ethanol-precipitated, rRNA-depleted RNA pellet was resuspended in 18 µL of the Fragment, Prime, Finish Mix from Illumina's TruSeq Stranded mRNA LT product (RS-122-2101). RNA was fragmented for 1 min. at 94°C. Subsequently, cDNA copies of the RNA were synthesized using SuperScript II Reverse Trancriptase (Invitrogen 18064022) and double-stranded cDNA was purified with Agencourt AMPure XP beads (Beckman Coulter A63880) following Illumina's TruSeq protocol (RS-122-9004DOC). The 3' ends of the cDNA were adenylated, indexing adapters were ligated onto the ends, then the cDNA was amplified by PCR using primers that anneal to the ends of the adapters. The Pippin Blue DNA size selection system was used to select for fragments <500 bp.