GSM7814517: before starvation, biol repl 2; Nakaseomyces glabratus; RNA-Seq
Briefly, 5 mL of culture was added directly into 7.5 mL of pre-chilled methanol (−50°C) and incubated in an ethanol-dry ice bath at that temperature for at least 20 min. When all samples were in the ethanol-dry ice bath for > 20 minutes, cells were collected by centrifugation and quickly washed with ice-cold water to remove the methanol and resuspended in RNAlater solution (Qiagen, 76104) for at least 2 hours. Cells were centrifuged to remove the RNAlater, flash-frozen in liquid nitrogen and stored at -80 C until RNA-extraction. For each sample, ∼5x10^7 cells were collected and total RNA was extracted using a MasterPure Yeast RNA purification kit (Biosearch Technologies, MPY03100) following the manufacturer's protocol. RNA-seq libraries were prepared with the TruSeq RNA Library Preparation Kit v2 (Illumina) with the mRNA purification option.