title: GSM7903703: NuclearExtract_replicate2_stress_Nab2-wild type; Saccharomyces cerevisiae; RNA-Seq
library_name: GSM7903703
library_construction_protocol: 1) Whole cell extracts: Cells were harvested and snap-frozen in liquid N2. Pellets were resuspended in lysis buffer and further processed with acid-saturated phenol and chloroform to extract total RNA. Purified RNA was processed with Dynabeads with oligodT(25) for PolyA selection. 2) Nuclear extracts: Cells were spheroplasted with Zymolyase to remove cell walls. Nuclei were released in buffer containing 20% Ficoll and dounced to mechanically assist nucleus release. Nuclei were separated from remaining cell material/debris through sucrose gradient fractionation. The enriched nucleus fraction was resuspended in lysis buffer and further processed with acid-saturated phenol and chloroform to extract total RNA. Purified RNA was processed with Dynabeads with oligodT(25) for PolyA selection. The eluted poly(A) RNA was used to generate strand-specific sequencing libraries using the NEXTflex Rapid Directional RNA-Seq Library Prep Kit (BioO)