Published 2016
| Version v1
Dataset
Open
Lp02_2nd
Description
2nd time collection: Jan 2016. Defocus -6.Lp02 is thymidine auxotroph of the parental strain Lp01 originating from Philadelphia-1 strain. Cells were grown in AYE liquid culture with following supplements: ferric nitrate, cysteine hydrochloride and thymidine. Scrape cells off a plate, dissolve in the media, adjust the cell density to 0.6 and make a dilution series. Start O/N culture with all of them and freeze cells next day from the one in early stationary phase that is OD ~ 3.0
Files
keyimg_dga2016-01-15-20.jpg
Technical info
Tilt Series Date: 2016-01-15
Data Taken By: Debnath Ghosal Species / Specimen: Legionella pneumophila Strain: Philadelphia-1_Lp02 Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.50°, constant angular increment, dosage: 100.0 eV/Ų, defocus: -6.0 μm, magnification: 26000x. Microscope: Janelia Titan Acquisition Software: UCSFTomo Upload Method: pipeline Processing Software Used: Raptor Collaborators and Roles: Strain sent to us by Joseph P. Vogel, Washington University School of Medicine. Data collected and processed by Debnath Ghosal. This data was collected at the Janelia Krios. Purification / Growth Conditions / Treatment: Lp02 is thymidine auxotroph of the parental strain Lp01 originating from Philadelphia-1 strain. Cells were grown in AYE liquid culture with following supplements: ferric nitrate, cysteine hydrochloride and thymidine. Scrape cells off a plate, dissolve in the media, adjust the cell density to 0.6 and make a dilution series. Start O/N culture with all of them and freeze cells next day from the one in early stationary phase that is OD ~ 3.0. Added gold before freezing. Sample Preparation: Grids were stored in Janelia from previous imaging. See previous Lp02 sample prep. Vitrobot (MK iv) settings: 100% humidity, 4µl sample, Blot force 6, blot time 3 sec. Wait time 0 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200. Add gold to cells prior to freezing: 1ul of gold to 4 ul of cells, freeze.External Files
Files available via S3 at https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-01-15-20/3dimage_72825/
tomo_area1000100010010_full.rec, Reconstruction (Pixel Size 1.68 nm), 430.6 MB Download tomo_area1000100010010.mrc, Tilt Series (Pixel Size 0.42 nm), 2.2 GB DownloadAdditional details
- DOI
- 10.22002/b3k1y-pc559
- TILT_SERIES_ID
- dga2016-01-15-20
- NIH
- HHMI
- Beckman Institute
- Gordon and Betty Moore Foundation
- Agouron Institute
- John Templeton Foundation
- Collected
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2016-01-15
- Created
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2016-01-21
- Updated
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2016-01-22