Hi Marco, An alternative would be to scrap the SeMet crystals and collect anomalous data on native crystals at I23, Diamond Light Source. We are optimised for S-SAD and could likely help you with your phasing. Happy to talk off-list if you'd like to come and do an experiment (I am a beamline scientist on I23), just drop me and email (christian....@diamond.ac.uk). If you'd like to check the feasibility of the experiment, I made a web app where you put in a bit of info and it will tell you the likelihood of successful S-SAD phasing on I23 - https://diamondi23.anvil.app/.
Best, Chris ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of dbellini <dbell...@mrc-lmb.cam.ac.uk> Sent: Tuesday, May 14, 2024 06:23 To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> Subject: Re: [ccp4bb] Experimental phasing Selenomethionine data collection etc. tips [You don't often get email from dbell...@mrc-lmb.cam.ac.uk. Learn why this is important at https://aka.ms/LearnAboutSenderIdentification ] Hi Marco, A few suggestions that I like to follow for MAD experiments: Before everything, check you have at least about 1 SeMet per 100 residues Then before crystallisation check by MassSpec that SeMet is properly incorporated in your protein After crystallisation collect first on the peak with (very) high redundancy and as little/gentle dose as possible Collecting the other wavelengths should give you better starting phases/maps, which might be very helpful at your resolution of 2.8 (especially if it is a very anisotropic 2.8...) Automated pipelines are so good nowadays, if you collect good data they should solve it without problems (as long as your crystal is not suffering from other pathologies like twinning or pseudosymmtries). Good luck! D On 2024-05-14 01:17, Marco Bravo wrote: > CAUTION: This email originated from outside of the LMB: > .-owner-ccp...@jiscmail.ac.uk-. > Do not click links or open attachments unless you recognize the sender > and know the content is safe. > If you think this is a phishing email, please forward it to > phish...@mrc-lmb.cam.ac.uk > -- > > Hello all, > I have a data collection trip next week and plan to collect data on > selenomethionine derivative crystals at the al831 beamline. Are there > any resources, tips, tutorials, literature etc. That you can recommend > to help me prepare for these experiments. Also is there a way to plug > in the experimental data into ccp4 cloud to do the automatic structure > solution? Do I need native and derivative data to solve the structure? > Last trip I collected a seemingly 2.8 angstrom resolution data on a > crystal of the native protein but could not get a solution depsite > extensive molecular replacement attempts. 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