Agro_AD1231

Description

Technical info

Tilt Series Date: 2015-08-03

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: 155.0 eV/Ų, defocus: -8.0 μm, magnification: 22500x.

Microscope: Caltech Polara

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.

Purification / Growth Conditions / Treatment: Grow culture in AB medium (pH 7) at 30C. For induction, use 375 µl of a decent ON culture to inoculate 5 ml AB mes pH 5.8 medium containing 100 µM Acetosyringone and antibiotic (Cb 50). Grow at 20C for 18 hr. Concentrate cells 5 times by centrifuging. Freeze cells.

Sample Preparation: Vitrobot (MK III, MK IV was not working) settings: 100% humidity, 4µl sample, 5% BSA treated gold. Blot offset -5, blot time 4 sec. Wait time 0 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200.

Attached Files

20150803_1231_1.mrc, Tilt Series (Pixel Size 0.48 nm), 4.3 MB