E_coli_EPEC_8612

Description

Technical info

Tilt Series Date: 2020-02-13

Data Taken By: Mohammed Kaplan

Species / Specimen: Escherichia coli

Strain: 8612, EPEC Δrof1-escLΔrof3-mpcΔescN-espFΔflhBAΔfliOPQRΔminCD

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 3°, constant angular increment, dosage: 105.0 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: These cells were obtained from Ritesh (Ilan Rosenshine lab) and Amit (Sigal ben Yehuda lab, . both from the Hebrew university). They were present during sample preparation.

Purification / Growth Conditions / Treatment: An overnight culture from -80C stock 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: The samples were hold with a tweezer from the starvation plates and washed with PBS. Blotted . with different conditions (B

Attached Files

EPEC_8612_025_af.mrc, Tilt Series (Pixel Size 0.449 nm), 1.9 GB

EPEC_8612_025_af_part41_2.rec, Reconstruction (Pixel Size 1.796 nm), 737.3 MB