title: RNA-seq analysis of fission yeast WT, and cbf11DBM and Pcut6MUT mutants in standard and ammonium-supplemented complex media, and upon cerulenin treatment
design: RNA-seq analysis of fission yeast WT, and cbf11DBM and Pcut6MUT mutants in standard and ammonium-supplemented complex media, and upon cerulenin treatment
library_construction_protocol: Samples were prepared from 3 biological replicates. Two separate experiments were performed: 1) Cells were cultured to exponential phase in the complex YES medium with or without ammonium supplementation. 2) Cells were cultured to exponential phase in the complex YES medium and then treated with DMSO (control) or 20 Î¼M cerulenin for 1 hour. For each sample, 10 mL were harvested (600 g, 2 min) and the cell pellet was flash-frozen. Total RNA was isolated using a hot acidic phenol method followed by phenol-chloroform extractions and precipitation (PMID: 12854975). Extracted RNA was treated with TURBO DNase (Thermo Fisher Scientific) and purified using Qiagen RNeasy columns. RNA quality was assessed on Agilent Bioanalyzer. The sequencing libraries were synthesized using KAPA mRNA HyperPrep Kit (Illumina platform) (Roche, KK8581).