title: GSM1289257: Cerevisiae_RPF; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: mRNA-seq: Total RNA was subjected to rRNA depletion, randomly fragmented by partial alkaline hydrolysis and then size-selected RNA fragments (27-40 nt) were used for library preparation. Ribosome profiling: Cell extracts were processed exactly as described in Guo et al., 2010 (GSE22004). Cell extracts were digested with RNase I for 30 min at room temperature, and monosomes were purified by sucrose gradient centrifugation. Size-selected RNA fragments (~27-33 nt) were used for library preparation. Libraries were constructed exactly as described in Guo et al., 2010 (GSE22004). Libraries were prepared as in Grimson et al, 2008 (GSE12578) but with the following modifications. Because RNase I-digestion and alkaline-fragmentation products terminate with a 5'-hydroxyl and a 3'-phosphate, they were 3'-dephosphorylated before ligation to the 3' adaptor. Gel-purified 3'-ligation products were then 5'-phosphorylated before the 5'-ligation step.
attributes.tag: GEO Accession
attributes.value: GSM1289257