Agro_AD1484

Description

Technical info

Tilt Series Date: 2015-10-06

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: 160.0 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 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.

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.

dga2015-10-06-17_slicer16257.jpg: stackin gon membrane dga2015-10-06-17_slicer16256.jpg: stackin gon membrane

Attached Files

AD1584_B1tgt0017_full.rec, Reconstruction (Pixel Size 1.92 nm), 445.4 MB

AD1584_B1tgt0017.mrc, Tilt Series (Pixel Size 0.48 nm), 3.4 GB