title: GSM2640803: Yeast Total RNA replicate 2; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: The isolation of SG cores was adapted from Jain et al., 2016 and Wheeler et al. 2017.  Stress granules were purified through a series of differential centrigutation step, bound to ani-GFP antibody, and purified by binding to Protein A dynabeads. The beads were resuspended in a Protease K solution and incubated for 15 mins at 37°C.  Trizol LS reagent was added to the samples and RNA was extracted following the manufacturers protocol. Following isopropanol precipitation, the RNA pellet was re-suspended in 20 µL RNase-free H2O.  RNA was then Ribzeroed and DNase treated. Yeast sample: RNA was phenol:chloroform extracted, Ribzeroed, and DNase treated cDNA libraries were prepared following manufacturer’s protocol from three replicates of 10 ng ribosomal-depleted RNA from cells and SG cores using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina Yeast samples: Libraries were constructed using the Bio Scientific NEXTflex Rapid Illumina Directional RNA-Seq Library Prep Kit.
attributes.tag: GEO Accession
attributes.value: GSM2640803