alias: GSM1919477
title: GSM1919477: Membrane ribosomes, CHX; Saccharomyces cerevisiae; OTHER
library_construction_protocol: Cells were lysed using a cryogenic mixer mill. Soluble and membrane-bound ribosomal fractions were separated by centrifugation. SRP72-TAP was immunoprecipitated and copurifying ribosomes were recovered. Polysomes and monosomes were separated using sucrose gradient ultracentrifugation. Samples were then digested with RNase I, and ribosomes were isolated by ultracentrifugation through a sucrose cushion. RNA was extracted from the ribosomal pellet. RNA fragments of approximately 30 nucleotides were extracted from a denaturing polyacrylamide gel. A DNA adapter (CTGTAGGCACCATCAAT) was ligated to the 3' of fragments and used to prime reverse transcription. cDNA were circularized and rRNA-originating fragments were removed using biotinylated subtraction oligonucleotides. Libraries were barcoded during PCR amplification.
identifiers.namespace: GEO
identifiers.id: GSM1919477