Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Cells were resuspended in 0.5 ml AE buffer with 10% SDS, 0.5 ml phenol, pH 5.3, and approximately 0.3 ml glass beads. Samples were vortexed 30 min at 4°, incubated 10 min at 65°, frozen at -80°, and spun 10 min in microcentrifuge at room temperature. The aqueous layer was extracted twice with phenol/CHCl3, and the RNA was precipitated with ethanol. 100 ug RNA was treated with DNase I for 10 min at room temp, applied to RNeasy spin column (Qiagen), treated with DNase I on the column, and eluted in water. RNA libraries were prepared for sequencing using standard Illumina protocols at the Duke University Sequencing core facility