Illumina HiSeq 2000 sequencing; Phenotypic analysis of the fil1 transcription factor from S. pombe
Phenotypic analysis of the fil1 transcription factor from S. pombe
Cells were grown at 32C in EMM2 (Moreno et al. Methods Enzymology 194: 795). 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).