Vibrio cholerae SAD2090

Description

Technical info

Tilt Series Date: 2020-09-09

Data Taken By: Stefan Kreida

Species / Specimen: Vibrio cholerae

Strain: SAD2090

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 2°, constant angular increment, dosage: 150.0 eV/Ų, defocus: -8.7 μm, magnification: 26000x.

Microscope: Caltech Titan Krios

Acquisition Software: Serial EM

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Strain from Ankur B Dalia, ankdalia@indiana.edu, PhD, Assistant Professor, Indiana University. . Cell growth & preparation, data collection and frame alignment by Stefan Kreida.

Purification / Growth Conditions / Treatment: 100 uL O/N preculture used to inoculate 3 mL LB, 20 mM MgSO4, 10 mM CaCl2, 100 uM IPTG. . Cells were grown for 7 h (30C) on 170 rpm. 100 uL culture was then spun down at 5,000 x g (5') . and resuspended in 100 uL aquarium salt (Instant Ocean), 7 g/L.

Sample Preparation: BSA/10 nm gold suspension: 1 mL 10 nm gold suspension mixed with 0.25 mL 5% BSA, spun . at 14,000 rpm (10'). Supernatant removed so that 35 uL remained, which was used to . resuspend the gold particles. 30 uL cells mixed with 5 uL gold suspension. 3 uL sample . applied in Vitrobot at 100% humidity. Grid: Quantifoil R2/2, Cu NH2, LotID 186865.

Attached Files

G12_SAD2090_TS13_ali.mrc, Tilt Series (Pixel Size 0.33 nm), 2.9 GB

G12_SAD2090_TS13_ali_part61_2.rec, Reconstruction (Pixel Size 1.32 nm), 737.3 MB