DOI: 10.22002/jz20f-xek09
title: Agro_AD1408_from plate_20150831

description: Agro_AD1408_GFP-VirB11 (Kan+AS plates, Mes pH5.8). Add 400 ul of induction media to one of the induction plates (containing lawn of Agrobacterium AD1408) and wash the plate by gently rolling the liquid. I had 20ug/ml Kan to keep the cells happier. Add gold to this: 1ul of gold to 4 ul of cells and freeze.
description: <strong>Tilt Series Date:</strong> 2015-08-31</p>
<strong>Data Taken By:</strong> Debnath Ghosal</p>
<strong>Species / Specimen:</strong> Agrobacterium tumefaciens</p>
<strong>Strain:</strong> </p>
<strong>Tilt Series Settings:</strong>  Single Axis, tilt range: (-50.0°, 50.0°), step: 1.0°, constant angular increment, dosage: 170.0 eV/Å²,  defocus: -10.0 μm,  magnification: 22500x. </p>
<strong>Microscope:</strong> Caltech Polara</p>
<strong>Acquisition Software:</strong> UCSFTomo</p>
<strong>Upload Method:</strong> pipeline</p>
<strong>Processing Software Used:</strong> UCSFTomo</p>
<strong>Collaborators and Roles:</strong> Strain constructed by Anath Das. Data collected and processed by Debnath Ghosal using Caltech tomography pipeline. This data was collected at the Janelia Krios.</p>
<strong>Purification / Growth Conditions / Treatment:</strong> Grow AD1408 cells on AB solid medium (pH 7) at 30C. For induction, use one single colony and spread on induction plates (AB Mes pH 5.8) containing Kan (20ug/ml)+AS (200µM). Grow at 19C for 48 hr. Add 400 ul of induction media to one of the induction plates (containing lawn of Agrobacterium AD1408) and wash the plate by gently rolling the liquid. Add gold to this: 1ul of gold to 4 ul of cells, freeze.</p>
<strong>Sample Preparation:</strong> 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.</p>
description: Files available via S3 at https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2015-08-31-13/3dimage_61040/</p>AD1408_plate0010_full.rec,  Reconstruction (Pixel Size 1.92 nm), 445.4 MB  
        
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        AD1408_plate0010.mrc,  Tilt Series (Pixel Size 0.48 nm), 2.9 GB  
        
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        dga2015-08-31-13_slicer18050.png, , 792.5 kB  
        
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        dga2015-08-31-13_slicer18051.png, , 791.8 kB  
        
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        dga2015-08-31-13_slicer18052.png, , 787.4 kB  
        
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