RELPDBePDBePDBe2016-04-142016-05-182016-06-152016-06-22Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessmentJoseph APMalhotra SBurnley TWood CClare DKWinn MTopf MCryo-EM, Refinement of atomic models, assessmentJoseph APMalhotra SBurnley TWood CClare DKWinn MTopf MRefinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessmentMethods1004249201626988127doi:10.1016/j.ymeth.2016.03.007GroELGroELtetradecamer10.0560.056GroELEscherichia colicytoplasm0.80.8tetradecamerfalseP0A6F5GO:1990220IPR001844singleParticleparticle47.450 mM Tris-HCl, 50 mM KCl and 10 mM MgCl2NEGATIVE3.5ul of protein was added to glow discharged C-flat r2/2 grid which were then blotted and plunge frozen400 mesh C-flat r2/2ETHANE40100HOMEMADE PLUNGER3.5ul of protein was added to glow discharged C-flat r2/2 grid which were then blotted and plunge frozenFEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN3002.71.02.8130000.047400.0FEI TITAN KRIOS AUTOGRID HOLDER909190Objective lens astigmatism was corrected at the magnification used to collect the dataGatan0.020.0The images were recorded using a dose rate of ~8 electrons/A2/s (~9 electrons/pixel/s) with 20 movie frames collected over a 4 second exposure2015-10-09GATAN K2 QUANTUM (4k x 4k)537032Each image is summation of 20 frames recorded over 4 seconds32.GroEL was aligned and reconstructed in RelionEach particleC7OTHER3.26OTHERRelion, Ctffind317400emd_3407_fsc.xml