title: Illumina HiSeq 2500 sequencing; Ribosome profiling of budding yeast Saccharomyces cerevisiae in eEF3 depleted conditions
design: Ribosome profiling of budding yeast Saccharomyces cerevisiae in eEF3 depleted conditions
library_construction_protocol: Cells were collected by rapid vacuum filtration and subsequent flash-freezing in liquid nitrogen and cryogenic milling. Cultured at 30 degrees Celsius in 700 ml of synthetic drop-out SC -met-cys medium supplemented with or without 0.5 mM of methionine. TRI Reagent (AM9738, ThermoFisher) was used to extract the RNA, followed by clean-up using RNA Clean & Concentrator -25 Kit (Zymo Research).  RNA-Seq libraries were constructed using ScriptSeq Complete Gold Yeast Kit (Epicentre). Ribo-Seq libraries were prepared essentially as per Ingolia and colleagues, NatMeth 2012 (PMID: 22836135) with modifications on rRNA removal (Ribo-Zero Gold rRNA Removal Kit -Yeast) and sample purification procedures between enzymatic steps (RNA Clean & Concentrator -5 Kit).
attributes.tag: Experimental Factor: compound
attributes.value: methionine
attributes.tag: Experimental Factor: dose
attributes.value: 0.5
attributes.tag: Experimental Factor: fraction
attributes.value: rRNA-depleted total RNA