JV3559_Janelia

Legionella thyA ΔdotG.This is a single mutant that lacks DotG.<strong>Tilt Series Date:</strong> 2017-01-08</p> <strong>Data Taken By:</strong> Debnath Ghosal</p> <strong>Species / Specimen:</strong> Legionella pneumophila</p> <strong>Strain:</strong> Philadelphia-1_Lp02</p> <strong>Tilt Series Settings:</strong> Single Axis, tilt range: (-60.0°, 60.0°), step: 1.50°, constant angular increment, dosage: 100.0 eV/Ų, defocus: -6.0 μm, magnification: 26000x. </p> <strong>Microscope:</strong> Janelia Titan</p> <strong>Acquisition Software:</strong> UCSFTomo</p> <strong>Upload Method:</strong> pipeline</p> <strong>Processing Software Used:</strong> Raptor</p> <strong>Collaborators and Roles:</strong> Strain sent to us by Joseph P. Vogel, Washington University School of Medicine. Data collected and processed by Debnath Ghosal at F30, Caltech.</p> <strong>Purification / Growth Conditions / Treatment:</strong> Cells were grown in AYE liquid culture with following supplements: ferric nitrate, cysteine hydrochloride and thymidine. Scrape cells off a plate, dissolve in the media, adjust the cell density to 0.6 and make a dilution series. Start O/N culture with all of them and freeze cells next day from the one in early stationary phase that is OD ~ 3.0. This time cell density was ~3.4</p> <strong>Sample Preparation:</strong> 8 grids were frozen for the JV3559 strain at OD~ 3.4, 4 grids at density x and another 4 at density 2x. Vitrobot (MK iv) settings: 100% humidity, 4µl sample, Blot force 6, blot time 3 sec. Wait time 0 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200. Add gold to cells prior to freezing: 1ul of gold to 4 ul of cells, freeze.</p>Files available via S3 at https://renc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2017-01-08-54</p>JV3559_20170106_area10032_full.rec, Reconstruction (Pixel Size 1.68 nm), 445.4 MB <a role="button" class="ui compact mini button" href="https://renc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2017-01-08-54/3dimage_80380/JV3559_20170106_area10032_full.rec" > <i class="download icon"></i> Download </a></p> JV3559_20170106_area10032.mrc, Tilt Series (Pixel Size 0.42 nm), 2.3 GB <a role="button" class="ui compact mini button" href="https://renc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2017-01-08-54/rawdata/JV3559_20170106_area10032.mrc" > <i class="download icon"></i> Download </a></p&gt