A413T co-overexpression
Contributors
Researcher:
Description
Technical info
Tilt Series Date: 2013-09-11
Data Taken By: Ariane Briegel
Species / Specimen: Escherichia coli
Strain: RP 437
Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1°, constant angular increment, dosage: 160.0 eV/Ų, defocus: -8.0 μm, magnification: 41000x.
Microscope: Caltech Polara
Acquisition Software: UCSFTomo
Upload Method: reupload_downloaded
Processing Software Used: Raptor
Collaborators and Roles: Laura Kiessling
Purification / Growth Conditions / Treatment: Grew UU1607/pJC3/pPM25 overnight in 5mL TB + 3.8µl Cm + 5µl CarbDiluted back 1:50 into 200mL TB + 152µl Cm + 200µl Carb + 50µl 1M IPTG + 200µl Na-salicylateGrew ~2 hours to mid-log phaseSpun down 10' at 5,000 x gResuspended in 300µl lysis buffer + 0.1µl PMSF + 0.75 µl 1,10-phenanthrolineAdded 33.3µl (2mg/ml) fresh lysozyme solution (20mg/ml in 10mM Tris-HCl, pH 8.0)Incubated 45' at 37°CAdded 1.67µl 1M MgCl2, 0.2µl 1M CaCl2, 0.33µl DNase IIncubated 30' at 37°CSpun down 4000 g 5 min Resuspended in activity buffer (cold room) without glycerol
Attached Files
A413Tatlas2zerodegreemin0007.mrc, Tilt Series (Pixel Size 0.26 nm), 3.4 GB
A413Tatlas2zerodegreemin0007.rec, Reconstruction (Pixel Size 0.0 nm), 2.8 GB
Files
keyimg_aba2013-09-11-3.jpg
Additional details
Identifiers
- DOI
- 10.22002/8djjd-djs17
- TILT_SERIES_ID
- aba2013-09-11-3
Funding
- NIH
- HHMI
- Beckman Institute
- Gordon and Betty Moore Foundation
- Agouron Institute
- John Templeton Foundation
Dates
- Collected
-
2013-09-11
- Created
-
2013-09-11
- Updated
-
2013-10-08