Pseudomonas_aeruginosa_DfliP

Description

Technical info

Tilt Series Date: 2019-05-25

Data Taken By: Mohammed Kaplan

Species / Specimen: Pseudomonas aeruginosa

Strain:

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 2°, constant angular increment, dosage: 120.0 eV/Ų, defocus: -8.0 μm, magnification: 19500x.

Microscope: Caltech Titan Krios

Acquisition Software: Serial EM

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: I obtained these cells from Dianne Newman lab from the transposon library (mutant 10239 from . the full library excel file). Shannon Park helped me to get the cells.

Purification / Growth Conditions / Treatment: Cells were first grown on LB plate with 100 ug/ml gentamicin over night (from -80 C stock). . Next dat, one colony was inoculated in 2 ml of LB (with 100 ug/ml gentamicin) and incubated . for 16 hours at 37 C with 175 rpm. After cells were mixed with gold beads (coated with BSA), . 4 ul were added to the grids, blotted and plunge frozen.

Sample Preparation: Different plunge-freezing parameters: 1) blot force 6, blot time 3s, drain time 1s. 2) blot force 3, . blot time 2s, drain time 1s. 3) blot force -5, blot time 4s, drain time 1s. 4) blot force -3, blot time . 2s, drain time 1s. All conditions were at room temperature with 100 % humidity.

Attached Files

P_aeruginosa_DfliP20.mrc, Tilt Series (Pixel Size 0.449 nm), 2.8 GB

P_aeruginosa_DfliP20_full.rec, Reconstruction (Pixel Size 1.796 nm), 737.3 MB

P_aeruginosa_DfliP20.mrc.mdoc, 31.2 kB