Pseudomonas aerugionsa PA103

Description

Technical info

Tilt Series Date: 2019-06-19

Data Taken By: Steven Wang

Species / Specimen: Pseudomonas aeruginosa

Strain:

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 95.0 eV/Ų, defocus: -6.0 μm, magnification: 19500x.

Microscope: Caltech Titan Krios

Acquisition Software: SerialEM

Upload Method: pipeline

Processing Software Used: Jensen Lab tomography pipeline (RAPTOR for alignment)

Collaborators and Roles: Alan Hauser lab from Northwestern University

Purification / Growth Conditions / Treatment: The cells were grown in LB with 50 ug/mL gentamicin. Incubate the cells for 1 hour at 37 C 200 rpm at a dilution factor of 50. Then add IPTG to reach a final concentration of 1 mM. Incubate for 6 hours at 37 C at 200 rpm. Then remove most of the filamentous cells by 3000xg centrifugation for 10 min. Collect the minicells from the supernatant with another centrifugation at 15,000xg for 30 min.

Sample Preparation: Mix 40 uL of cells with gold beads concentrated from 200 uL Au solution + 50 uL 5% BSA. Plunge freeze the samples with liquid ethane and propane with a blotting force of 4, a blotting time of 3 s or 4s and a draining time of 1 s.

Attached Files

Pa_cell_target_12.mrc, Tilt Series (Pixel Size 0.45 nm), 5.7 GB

Pa_cell_target_12_full.rec, Reconstruction (Pixel Size 1.8 nm), 589.8 MB