pk1-1-3

Description

Technical info

Tilt Series Date: 2019-07-21

Data Taken By: William Nicolas

Species / Specimen: Escherichia coli

Strain: PK1, BL21 dHisdHA RQABpCDF+EFGpRSF S2-ZC

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 3°, constant angular increment, dosage: 134.0 eV/Ų, defocus: -6.0 μm, magnification: 26000x.

Microscope: Caltech Titan Krios

Acquisition Software: Serial EM

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Petya Krasteva (supplied the OE lines)

Purification / Growth Conditions / Treatment: Grown from glycerol stock in TB medium with Kana (40ug/mL) and Strepto (70ug/mL) (220rpm - 37C) . until OD600 reaches 1. Induction of OE with IPTG 0.7mM and grown OE at 220rpm - 18C.

Sample Preparation: OD600 is checked (usually around 4/5 after ON induction) and adjusted to 0.4.. . Quantifoil Finder Cu NH2 R2/2 glowdischarged 15mA - 1min. . FM: 50uL BSA 5% + 5uL FM solution - Spin down 15000g - 15min -discard SN and resuspend FM pellet . with 40uL of bacterial culture. . Blotting conditions: 100% hum - 30C - manually backblotted 4 to 5secs. .

Attached Files

pk1-1-3_sq5_cell1.mrc, Tilt Series (Pixel Size 0.34 nm), 1.9 GB

pk1-1-3_sq5_cell1_full.rec, Reconstruction (Pixel Size 1.36 nm), 737.3 MB