H. pylori 26695 / 20 nm gold-DNA

Description

Technical info

Tilt Series Date: 2014-01-21

Data Taken By: Yi-Wei Chang

Species / Specimen: Helicobacter pylori

Strain: 26695

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 190.0 eV/Ų, defocus: -8.0 μm, magnification: 22500x.

Microscope: Caltech Polara

Acquisition Software: UCSFTomo

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: H.pylori 26695 from ATCC, prepared by Lee Rettberg

Purification / Growth Conditions / Treatment: H.pylori cells grown in BLBB + 10% heat inactivated fetal bovine serum at 37 degrees in microaerobic chamber with GasPak EZ Campy Sachets for 3 days, OD600=0.4. Spun cells down at 3000rpm for 5 minutes to concentrate before using; resuspended pellet.

Sample Preparation: Combined 20ul cells with 10ul 10 nm gold fiducials (100ul gold and 25ul BSA) and 10ul 20 nm streptavidin gold-biotin DNA. Grids frozen 7-20 minutes after combining DNA labeled gold with concentrated cells. Vitrobot settings: 100% humidity, 22.5 C, -3 offset, 4.5 sec blot time, Ethane/propane mix.

Attached Files

20140121_LR005_30003.mrc, Tilt Series (Pixel Size 0.48 nm), 3.4 GB

20140121_LR005_30003_full.rec, Reconstruction (Pixel Size 1.92 nm), 712.7 MB