title: GSM4658963: RNA-seq from untreated cells, rep 3; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: Samples were rapidly collected using filtration and ground to a fine powder using a Freezer/Mill (SPEX SamplePrep) and stored at -80 °C. To prepare RNA-seq libraries with fragment sizes that matched the ribosome profiling libraries, 25 µg of total RNA was poly(A)-selected, digested into 27–33 nt fragments, and used to prepare libraries in parallel with the ribosome-protected fragments. For ribosome profiling libraries, aliquots of lysate were digested with 1 U/μL RNase I (Ambion) for 30 min at room temperature and then run on a 10–50% sucrose gradient to purify monosomes (Subtelny et al., 2014). RNAs from both RNA-seq and ribosome profiling were then size-selected, ligated to adapters, reverse-transcribed, and amplified (Subtelny et al., 2014).
attributes.tag: GEO Accession
attributes.value: GSM4658963