GSM3185739
GSM3185739: Riboseq.replete.WT_1; Saccharomyces cerevisiae; OTHER
Cells were lysed by vortexing with glass beads in polysome lysis buffer (20 mM HEPES-KOH pH 7.4, 2 mM MgAc, 0.1 M KAc, 0.1 mg/mL cycloheximide, 1% TritonX-100). Extracts were clarified at 21,000 × g for 20 minutes. Lysate was digested with 15U/A260 RNase I. 80S footprints were isolated from a sucrose gradient, enriched for small RNAs, and size-selected from 26-31 nucleotides. A 3′ adapter was ligated and samples treated with subtractive hybridization oligos to deplete ribosomal RNA. The RNA was reverse transcribed, circularized, and PCR amplified.
GEO
GSM3185739
GEO Accession
GSM3185739