Agro_piliprep7_highmag_16.09.15

Description

Technical info

Tilt Series Date: 2015-09-16

Data Taken By: Debnath Ghosal

Species / Specimen: Agrobacterium tumefaciens

Strain:

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 120.0 eV/Ų, defocus: -10.0 μm, magnification: 34000x.

Microscope: Caltech Polara

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: Patch Tracking

Collaborators and Roles: Pili prep from induced strain (AD1484) was done by Anath Das. Data collected and processed by Debnath Ghosal using Caltech tomography pipeline. This data was collected at the F30, Caltech.

Purification / Growth Conditions / Treatment: For pili prep, following buffer was used: 20 mM Tris-HCl, pH 7.8, 100 mM NaCl and 0.5% deoxycholate.

Sample Preparation: Vitrobot (MK iv) settings: incubate at 100% humidity for at least 15 min, 4µl sample, blot force 6, blot time 3 sec. Wait time 30 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200. Did not add any gold. I assumed we will only do high resolution imaging. However, later we decided to do tomography and used patch tracking to process the data.

Attached Files

AD1484_piliprep70014.mrc, Tilt Series (Pixel Size 0.32 nm), 3.4 GB

AD1484_piliprep70014.rec, Reconstruction (Pixel Size 1.28 nm), 890.9 MB

dga2015-09-16-15_slicer16262.png, 701.1 kB