Name: wt.noN.noCHX.mrna.1_s
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: RANDOM
Layout: SINGLE
Construction protocol: Cells were grown at a temperature of 32C in EMM2 (Moreno et al. Methods Enzymology 194: 795) containing NH4Cl as a nitrogen source. Cells were washed with EMM containing no NH4Cl (EMM-N), resuspended in EMM-N, and grown at 32C for 60 minutes. Total RNA was extracted using a hot phenol extraction method as described in Lyne et al. (2003) BMC Genomics 4:27. Clean up and DNase treatment were performed on with Purelink RNA microcolumns (Life Technologies) following the manufacturer’s instructions. Samples were ribo-depleted using Ribo-Zero Gold rRNA Removal Kit Yeast (Illumina) with a 4 ug input. 30 ng of ribo-depleted RNA was used as starting material for the NEXTflex Rapid Directional qRNA-Seq Kit (Bioo Scientific).