GSM3453495: tma20_sd_rna; Saccharomyces cerevisiae; RNA-Seq
Cells were resuspended in polysome lysis bufer (20 mM Tris pH 8.0, 140 mM KCl, 1.5 mM MgCl2, 100  g/ml cycloheximide, 1% Triton), flash frozen in liquid nitrogen and homogenyzed by grinding. Part of the cell lysates was used for mRNA isolation, another part was treated with ribonuclease I for polysome disassembly and applied to sucrose gradient fractionation with subsequent isolation of ribosome-bound RNA fragments from monosome fraction. mRNA was isolated using Oligo(dT) beads  and ribosome-bound RNA was isolated using acidic-phenol extraction. cDNA libraries for Ribo-Seq and RNA-Seq were constructed in parallel using standard ribosomal profiling library construction protocol (described in Ingolia NT et al., 2009, Science).
GEO
GSM3453495