GSM9416803: WT grown in7.35 mM potassium, biol rep 3; Saccharomyces cerevisiae; RNA-Seq
0.5 OD of cells was collected onto a 0.45 µm nylon filter (HNWP02500, Millipore Sigma) using a vacuum filtration system. Cell were resuspended in 200 µL of Lysis buffer (10 mM Tris [pH 8.0], 0.5% SDS, 10 mM EDTA). After adding of 200 µL acid phenol (pH 4.3), samples were vortexed for 30 s. Samples were then incubated at 65°C for 30 min in a thermomixer with intermittent shaking (2,000 rpm for 1 min every 15 min). Finally, a total of 400 µL of ethanol was added and the RNA was purified using the Direct-zol RNA Miniprep Plus kit (R2071; Zymo Research) according to the manufacturer's instructions, including the optional DNase digestion step. RNA integrity was confirmed using an Agilent Bioanalyzer. The Poly A selection + NEB Ultra II Directional RNA Library Prep Kit (New England BioLabs Inc., Massachusetts, USA) was used to generate cDNA and libraries according to the manufacturer's protocol. The libraries were indexed using Admera's in-house index primers and cleaned up using AFTMag NGS DNA Clean Beads (Abclonal science, RK20257). The indexed libraries were pooled and sequenced on the Illumina NovaSeq X plus platform with a read length of 2x150bp.