EMD-2290

Single-particle
14.0 Å
EMD-2290 Deposition: 21/01/2013
Map released: 13/03/2013
Last modified: 24/07/2013
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-2290

First 3D model of wild type MjHSP16.5 at 60 degree Celsius by CryoEM.

EMD-2290

Single-particle
14.0 Å
EMD-2290 Deposition: 21/01/2013
Map released: 13/03/2013
Last modified: 24/07/2013
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Method: Single-particle
Aggregation State: Particle
Specimen preparation [1]
Concentration: 0.8 mg/mL
Buffer
pH: 7.4
Details: 10mM HEPES, 100mM NaCl.
Grid
Details: GIG holey grids (LifeTrust, China) were treated with a glow discharge machine (Master Plasmer)
Vitrification
Cryogen name: ETHANE
Chamber temperature: 9 K
Instrument: FEI VITROBOT MARK IV
Method: The samples were first heated to 60 degree Celsius in the boiler, then 3.5 microlitre samples were added to the grid and blotted for 1.5 s with blot force 2 at 100% humidity;
Microscopy [1]
Microscope: FEI TITAN KRIOS
Illumination mode: FLOOD BEAM
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 300 kV
Nominal CS: 2.7 mm
Nominal defocus: 2.0 µm - 3.0 µm
Nominal magnification: 96000.0
Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Specimen holder details: Liquid nitrogen cooled
Temperature
Average: 85 K
Image Recording [1]
Detector category: CCD
Detector model: GATAN ULTRASCAN 4000 (4k x 4k)
Number of real images: 2119
Average electron dose per image: 20 e/Å2
Bits per pixel: 32.0
Details: Electron micrograph exposures were made with the automatic collection package Leginon.
Image processing [1]
Details: The particles were selected using an automatic selection program Gautomatch developed in Fei Sun lab (to be published). Octahedron symmetry were imposed during 3D reconstructing.
Final reconstruction
Resolution: 14.0 Å ( BY AUTHOR)
Resolution method: FSC 0.5 CUT-OFF
Number of images used: 4842
Algorithm: OTHER
Details: The final reconstructed density map was further sharpened by application of an amplitude correction algorithm in the program BFACTOR.
Applied Symmetry
Point group: O
Software [1]
Name Version Details
EMAN1 - -
Final 2D classification
Number of classes: 193
CTF correction
Details:Each image
Map
Format: CCP4
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: First model for reconstruction of wild type MjHSP16.5 at 60 degree Celsius
Details: ::::EMDATABANK.org::::EMD-2290::::
Geometry
X Y Z
Dimensions 256 256 256
Origin -128 -128 -128
Spacing 256 256 256
Voxel size 0.933 Å 0.933 Å 0.933 Å
Contour list
Primary Level Source
True 1.73 AUTHOR