EMD-5440 Experiments and Validation

Regulation of mammalian transcription by Gdown1 through a novel steric crosstalk revealed by cryo-EM

Single particle reconstruction
Overview of EMD-5440
Sample name: Bovine RNA polymerase II elongation complex
Organism: Bos taurus
Related EM entries by publication: EMD-5443, EMD-5442, EMD-5441

Map parameters

Minimum density: -0
Maximum density: 0.01
Average density: 0
Standard deviation: 0
Recommended contour level: 0 (emdb)

Sample information

Sample name: Bovine RNA polymerase II elongation complex
Protein: RNA polymerase II elongation complex

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: Spider, EMAN2
Number of particles: 24000
Number of class averages: 128
CTF correction: Each particle

Imaging

Session
Microscope model: JEOL 2100F
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 20.0
Holder model: GATAN LIQUID NITROGEN
Acquisition
Number of images: 150
Scanner model: ZEISS SCAI

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 1
Buffer: 50 mM Tris-Cl, 200 mM KOAc, 5mM MgCl2, 5 mM DDT
Specimen support: 200 mesh quantifoil grid with thin carbon layer
Vitrification
Apparatus: HOMEMADE PLUNGER
Cryogen: ETHANE
Humidity: 80
Protocol: Blot before plunging