Summary: | Exposure 1-1.3s Tilt Series Date: 2020-09-29 Data Taken By: William Nicolas Species / Specimen: Escherichia coli Strain: PK2, BL21 dHisdHA RQABpCDF+EFGpRSF Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 2°, constant angular increment, dosage: 56.0 eV/Ų, defocus: -8.0 μm, magnification: 11000x. Microscope: IECB Talos Artica Acquisition Software: Serial EM Upload Method: pipeline Processing Software Used: Raptor Collaborators and Roles: Petya Krasteva (head of the SBB lab at the IECB, Bordeaux° and Wiem . Abidi (graduate student) did the preps and helped with acquisition Purification / Growth Conditions / Treatment: BL21 cells were transformed with QAB - pCDF plasmid and EFG - pRSF . plasmid to express the inner-membrane macro-complex. . Selection done with Kanamycin and Streptomycin. . Induction of expression ON done with IPTG. . Spheroplasts are generated from the BL21 non transformed cells cells. Spheroplasts are processed through a high-pressure homogenizer and . further purification by ultracentrifugation of the membrane . fractions. . Liposomes are then generated from the membrane fractions by . processing through Potter and sonicator. Sample Preparation: Fiducials were prepared my method (see lab archive). . Grids: Quantifoil Cu NH2 R2/2. . Glowdischarged 3mA - 2min. BT3s - BF0. 4uL pipetted on the grids Files available via S3 at https://renc.osn.xsede.org/ini210004tommorrell/tomography_archive/wnj2020-09-29-27 lip10-2-sq3-tg18.mrc, Tilt Series (Pixel Size 0.39 nm), 1.7 GB Download lip10-2-sq3-tg18_full.rec, Reconstruction (Pixel Size 0.78 nm), 1.8 GB Download
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