DOI: 10.22002/60s5q-s5p09
title: Agro_AD1484

description: Agro_AD1484 (AD1484:A348dB/pAD2079,virB on a multicopy plasmid). Add 200 ul of induction media to one of the induction plate (containing lawn of Agrobacterium AD1484) and wash the plate by gently rolling the liquid. Take this liquid and add to the other plate. Repeat the same. This should get enough cells dissolved in the liquid.Add gold to this: 1ul of gold to 4 ul of cells.
description: <strong>Tilt Series Date:</strong> 2015-10-06</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: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 160.0 eV/Å²,  defocus: -8.0 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> Raptor</p>
<strong>Collaborators and Roles:</strong> Strain constructed by Anath Das. Data collected and processed by Debnath Ghosal using Caltech tomography pipeline</p>
<strong>Purification / Growth Conditions / Treatment:</strong> Grow AD1484 cells on AB solid medium with Cb (pH 7) at 30C. For induction, use one single colony and spread on induction plates (AB Mes pH 5.8) containing Cb (50ug/ml)+AS (200µM). Grow at 19C for 4days. Add 200 ul of induction media to one of the induction plate (containing lawn of Agrobacterium AD1484) 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/dga2015-10-06-18/3dimage_69997/</p>AD1584_B1tgt0018_full.rec,  Reconstruction (Pixel Size 1.92 nm), 445.4 MB  
        
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        AD1584_B1tgt0018.mrc,  Tilt Series (Pixel Size 0.48 nm), 3.4 GB  
        
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        dga2015-10-06-18_slicer19248.png,  ? .  -andrew, 326.8 kB  
        
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