E_coli_EPEC_8730

Description

Technical info

Tilt Series Date: 2020-02-26

Data Taken By: Mohammed Kaplan

Species / Specimen: Escherichia coli

Strain: 8730, EPEC 8612 + Δbfp

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 3°, constant angular increment, dosage: 115.0 eV/Ų, defocus: -6.0 μm, magnification: 19500x.

Microscope: Caltech Titan Krios

Acquisition Software: Serial EM

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Samples were prepared with Amit and Ritesh (from Ilan Rosenshine and Sigal ben Yehuda labs . at the Hebrew university).

Purification / Growth Conditions / Treatment: An overnight culture from a pre-prepared agar LB plate was made in 2ml LB and was grown at 37C in static conditions. The next day, 1 ml of cells were spun down at 4000 rpm for six minutes and concentrated 10 times. 3 ul of cells . were spotted on glow- discharged grids located on a DMEM high-glucose agar plates (1.5% agar) for 3 hours. Then, the grids were washed twice in PBS and transferred to a starvation-medium agar plates (Starvation medium:DMEM . without phenol red, vitamins and amino acids) and incubated for 1 hour at 37C. Subsequently, the samples were washed with PBS and plunge-frozen.

Sample Preparation: Samples were washed with PBS before blotting. Subsequently, the samples were automatically . blotted and plunge-frozen.

Attached Files

EPEC_8730_012.mrc, Tilt Series (Pixel Size 0.449 nm), 1.9 GB

EPEC_8730_012_part41_1.rec, Reconstruction (Pixel Size 1.796 nm), 737.3 MB