V. cholerae RT4236 (refreshed)

Description

Technical info

Tilt Series Date: 2015-12-19

Data Taken By: Yi-Wei Chang

Species / Specimen: Vibrio cholerae

Strain: O395-N1

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 160.0 eV/Ų, defocus: -6.0 μm, magnification: 27500x.

Microscope: Caltech Polara

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Strain from Ronald K. Taylor.

Purification / Growth Conditions / Treatment: V. cholerae 0395-N1 pMT5 grown in 5ml LB pH 6.5, 1 mM IPTG and 100 ug/mL ampicilin in glass tube shaking for overnight. After took the tube out from shaker, place the tube on bench for ~20 min, supernatant was removed and pellet was resuspended by gentle pipetting in 1ml fresh LB without IPTG nor ampicillin. Culture was incubated at room temperature for ~20 min before mixing with gold and freezing.

Sample Preparation: Vitrobot (MK iv) settings: 100% humidity, 4µl sample, 10 mg/ml BSA treated gold. Blot force 6, blot time 2.5 sec. Wait time 0 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200.

Attached Files

20151219_YWC206_10008_full.rec, Reconstruction (Pixel Size 1.56 nm), 534.5 MB

20151219_YWC206_10008.mrc, Tilt Series (Pixel Size 0.39 nm), 3.4 GB