GSM9139453: WT2_wt_high, LRY2992 (MATa SIR2 hmlΔ hmrΔ), nicotinic acid 400 μg/L; Kluyveromyces lactis; RNA-Seq
Ten OD₆₀₀ units of log-phase cells were washed in 500μl AE buffer (50mM NaoAc, ph=5.3; 10mM EDTA) and stored at -80°C. Frozen pellets were resuspended in 500μl AE/SDS buffer (10 parts AE: 1 part 10% SDS) and an equal amount of phenol-equilibrated with citrate (pH 4.5), vortexed and incubated at 65 °C for 4 min as per Schmitt et al., 1990. A second extraction with phenol/chloroform was followed by ethanol precipitation. Residual DNA was removed with DNase I (NEB M0303S). Poly(A)⁺ RNA selection and library preparation were completed by Azenta sequencing services. Paired-end 150 bp reads were generated on an Illumina NovaSeq 6000 sequencer.