GSM4103757: Sample157 [310]; Saccharomyces cerevisiae BY4741; RNA-Seq
For RNA extraction, wells growing at a relative growth rate close to 50% at the end of the third iteration of the automated re-inoculation culture were selected. RNA extraction was performed using the RiboPure RNA Purification Kit for yeast (Thermo Scientific, AM1926). The purity of extracted RNA was confirmed for selected samples using the Agilent RNA 6000 Nano Bioanalyzer Kit. For RNA extraction, wells growing at a relative growth rate close to 50% at the end of the third iteration of the automated re-inoculation culture were selected. RNA extraction was performed using the RiboPure RNA Purification Kit for yeast (Thermo Scientific, AM1926). The purity of extracted RNA was confirmed for selected samples using the Agilent RNA 6000 Nano Bioanalyzer Kit. The library was prepared as in (Bar-Ziv et al., 2016). In brief, the purified RNA was fragmented using the NEBNext® Magnesium RNA Fragmentation Module (New England Biolabs E6150S), poly(A)-selected using Dynabeads oligo-dT kit (Invitrogen, cat. No. 61012) and reverse-transcribed to cDNA using custom poly(T) primers barcoded for multiplexing as well as containing 4-nt-long unique molecular identifier. The resulting complementary DNA strands were pooled and purified (RNase H NEB M0297S, Agencourt AMPure XP Beckman Coulter A63881), a custom double stranded adapter ligated to the 3' end (Quick Ligation Kit, NEB M2200S), second cDNA strand synthesized (KAPAHiFi Hot-Start ReadyMix VWR 733-2430), amplified and 50 bp single-end sequenced on Illumina HiSeq 2500 using a primer complementary to the adaptor at the end opposite to the poly(A).