Rhodobacter1 558 lysed/PVP
Contributors
Researcher:
Description
Technical info
Tilt Series Date: 2013-10-07
Data Taken By: Ariane Briegel
Species / Specimen: Rhodobacter sphaeroides
Strain: ATCC 17023
Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1°, constant angular increment, dosage: 150.0 eV/Ų, defocus: -10.0 μm, magnification: 27500x.
Microscope: Caltech Polara
Acquisition Software: UCSFTomo
Upload Method: reupload_downloaded
Processing Software Used: SIRT
Collaborators and Roles: Judy Armitage
Purification / Growth Conditions / Treatment: Protocol:- grew 5mL cultures of 1558 (with kanamycin and IPTG) to log phase (OD600 between 0.12-0.15)- spun down 10' at 5000 xg- resuspended pellets in 2mL appropriate Buffer 1- Added 200µL fresh lysozme (50 mg/mL in 10mM Tris-HCl, pH 8.0)- Incubated 15' at RT- Added 11.2µL 1M MgCl2, 1.3µL 1M CaCl2, 2.2µL DNase I- Incubated 30' at 37C- Spun down 10' at 5000 xg- resuspended PVP in 80µL appropriate Buffer 2Buffer 1:Crowding agent1 g sucroseQ.S. 5mL 25mM Tris-HCl, pH 7.0Buffer 2:Crowding agentQ.S. 5mL 25mM Tris-HCl, pH 7.0
Attached Files
Rhodopvp27k0015.mrc, Tilt Series (Pixel Size 0.39 nm), 3.4 GB
Rhodopvp27k0015SIRTrot.rec, Reconstruction (Pixel Size 0.0 nm), 11.4 GB
Files
keyimg_aba2013-10-07-13.jpg
Additional details
Identifiers
- DOI
- 10.22002/xcath-4gk76
- TILT_SERIES_ID
- aba2013-10-07-13
Funding
- NIH
- HHMI
- Beckman Institute
- Gordon and Betty Moore Foundation
- Agouron Institute
- John Templeton Foundation
Dates
- Collected
-
2013-10-07
- Created
-
2013-10-07
- Updated
-
2013-10-09