title: NextSeq 500 sequencing; Phenotypic analysis of the fil1 transcription factor from S. pombe
design: Phenotypic analysis of the fil1 transcription factor from S. pombe
library_construction_protocol: Cells were grown at 32C in EMM2 (Moreno et al. Methods Enzymology 194: 795). Cells were grown in EMM2 (Moreno et al. Methods Enzymology 194: 795) in the presence of 10 mM 3-AT for 60 minutes at 32C.  This treatment mimics histidine starvation. 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).