Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 293 K / Instrument: LEICA EM GP / Details: 3 s blotting before plunging.
-
Electron microscopy
Microscope
FEI TITAN KRIOS
Specialist optics
Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recording
Film or detector model: GATAN K3 (6k x 4k) / Detector mode: SUPER-RESOLUTION / Number grids imaged: 3 / Number real images: 12233 / Average electron dose: 73.0 e/Å2 Details: The data from the three grids were also used to reconstruct the OCL-state US27-Gi-scFv16 complex (7RKY).
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Number selected: 12254340 Details: Including non-proteinous features. The actual number of intact complex particles was ~1,159,618.
CTF correction
Software - Name: cryoSPARC (ver. 3.1) Details: Final per-particle CTF values were determined by cryoSPARC Local CTF Refinement.
Startup model
Type of model: OTHER Details: Initial model created by cryoSPARC ab-initio reconstruction.
Final reconstruction
Resolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 3.1) Details: The resolution estimate was inflated due to the better-defined G protein region compared to the receptor region. Number images used: 268877
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