title: RNA-seq analysis of fission yeast mga2∆ and mga2∆ cbf11∆ mutants in standard and ammonium-supplemented complex media
design: RNA-seq analysis of fission yeast mga2∆ and mga2∆ cbf11∆ mutants in standard and ammonium-supplemented complex media
library_construction_protocol: Samples were prepared from 3 biological replicates of mga2Δ (MP815) and mga2Δcbf11Δ (MP836) cells. Cells were cultured to exponential phase in the complex YES medium (with or without ammonium supplementation), 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).