RELRCSBRCSBRCSB2015-10-052015-10-212015-10-212015-11-25Single-particle cryo-electron microscopy reconstruction of the eukaryotic large ribosomal subunitPassos DOLyumkis Dribosome, translationPassos DOLyumkis DSingle-Particle CryoEM Analysis at Near-Atomic Resolution from Several Thousand Asymmetric SubunitsJ.STRUCT.BIOL.192235244201526470814doi:10.1016/j.jsb.2015.10.0023jbsFULLOVERLAPSaccharomyces cerevisiae large 60S ribosomal subunitSaccharomyces cerevisiae large 60S ribosomal subunit12.560S large ribosomal subunitfalseSaccharomyces cerevisiaeBY4741yeastcytosol2.5ribosome-eukaryote: LSU 60SsingleParticleparticle6.850 mM HEPES-KOH, 100 mM KOAc, 5 mM MgOAc, 1 mM EDTA, 2 mM DTT400 mesh, 1.2x1.3 micron C-flat grids, plasma-treated for 5 secondsETHANE9077HOMEMADE PLUNGERA 3 uL sample was applied onto a freshly plasma-treated (6 seconds, Gatan Solarus plasma cleaner) holey carbon C-flat grid (Protochips, Inc.), allowing the sample to adsorb for 30 seconds. The sample was then plunge-frozen in liquid ethane using a manual cryo-plunger in ambient atmosphere at 4 degrees C.FEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN3002.70.52.522500.038167.0FEI TITAN KRIOS AUTOGRID HOLDERObjective lens astigmatism was corrected within the Leginon software.super-resolution imaging2014-09-02GATAN K2 (4k x 4k)183325each particleOTHER2.9OTHERFrealignB-factor of -50 applied to final map756534UJQHFLEXIBLE FITChimera, RosettaFSCREAL