We are excited to announce the launch of our cryo-recipes database: https://cryo-recipes.slac.stanford.edu, which is an assembly of parameters used for the structure determination of membrane proteins by cryoEM. We tried to capture everything from expression systems, biochemical purification conditions, over grid preparation, to imaging conditions and some image processing related numbers. It is currently unfortunately quite outdated, but we hope this is useful to the community, so please feel free to reach out for feedback or volunteer to submit some entries. The database is based on a github repository, so submission is welcome either via github or email.

To submit an entry, simply copy/paste below, edit and send an email to: cgati (at) stanford.edu

S2C2

Below the form for submission:

{
“grid_preparation”: {
“grid_protein_concentration”: 2,
“details”: “UltrAuFoil R 1.2/1.3 300”,
“comments”: “PEGylated gold grids”
},
“helices_tmd_oligomer”: 26,
“id”: 0,
“monomer”: 545,
“publication”: {
“emdb”: 3652,
“doi”: “10.1038/nmicrobiol.2017.70”,
“pdb”: “5NIK”,
“year_published”: 2017
},
“oligomer”: 1,
“type”: “ABC transporter”,
“affinity_tag_cleavage”: “-“,
“particle_size”: 545,
“imaging_conditions”: {
“frame_duration”: 0.8,
“total_dose”: 45,
“vpp”: “No”,
“symmetry_applied”: “C1”,
“resolution”: 3.3,
“super_resolution”: “Yes”,
“cs_corrector”: “No”,
“final_number_particles”: “91,162”,
“total_number_images”: 869,
“keV”: 300,
“detector”: “K2”,
“energy_filter”: “Yes”,
“total_exposure_time”: 16,
“pixel_size”: 1.36,
“software”: “RELION”
},
“native_source”: “Escherichia coli”,
“final_stabilizer”: “amphipol”,
“in_complex_with”: “-“,
“monomer_tmd_helices”: 26,
“name”: “-“,
“notes”: “-“,
“sample_preparation”: {
“affinity_tag_terminus”: “His-6, Flag (C)”,
“expression_organism”: “E. coli C43”,
“extraction_parameters”: “-“,
“composition_of_final_stabilizer”: “A8-35”,
“extraction_concentration”: “1.50%”,
“final_concentration_or_ratio”: “1:4”,
“initial_purification”: “HiTrap, Flag M2”,
“final_purification”: “Superose 6”,
“purification_details”: “first exchange to MNG”,
“extraction_method”: “DDM”,
“stabilizer_exchange”: “Yes”
},
“gene”: “MacAB-TolC”
},