Agro_AD1526_GFP-VirB11_20150826

Description

Technical info

Tilt Series Date: 2015-08-26

Data Taken By: Debnath Ghosal

Species / Specimen: Agrobacterium tumefaciens

Strain:

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 2.0°, constant angular increment, dosage: 160.0 eV/Ų, defocus: -8.0 μm, magnification: 26000x.

Microscope: Janelia Titan

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Strain constructed by Anath Das. Data collected and processed by Debnath Ghosal using Caltech tomography pipeline. This data was collected at the Janelia Krios.

Purification / Growth Conditions / Treatment: Grow AD1526 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 Cb (50ug/ml)+Kan (50ug/ml)+AS (200µM). Grow at 19C for 48 hr. Add 200 ul of induction media to one of the induction plate (containing lawn of Agrobacterium AD1526) and wash the plate by gently rolling the liquid. Add gold to this: 1ul of gold to 4 ul of cells, freeze.

Sample Preparation: 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.

dga2015-08-26-20_slicer14385.jpg: T6SS dga2015-08-26-20_slicer14394.jpg: Not visible in this plane

Attached Files

AD1526tomo0010_full.rec, Reconstruction (Pixel Size 1.68 nm), 430.6 MB

AD1526tomo0010.mrc, Tilt Series (Pixel Size 0.42 nm), 1.7 GB