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Sorry if I'm being stupid, but did you check them at room temperature? I know it sounds like something from pre-history! You say the crystals grow readily and you have some « big ones ». On a medium intensity (bending magnet) SR beamline, you will probably get between 10 and 20 images from a reasonable sized crystal (100 micron) cooled to 5 or 10 degrees C. Translate the crystal with a small beam to get extra "shots". If they diffract well at room temperature, have higher than monoclinic symmetry, you don't need super high resolution and you only need 1 wavelength to collect data (a lot of ifs) you could have solved your structure using only 1 or 2 crystals with less effort than solving your cryo problem. Cheers Andy Andy Thompson Synchrotron SOLEIL L'Orme de Merisiers BP 48 St Aubin 91192 Gif sur Yvette 0169359608 -----Message d'origine----- De : [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] De la part de Virgile Adam Envoyé : 09 April 2006 17:08 À : [email protected] Objet : Re: [ccp4bb]: cryo conditions *** For details on how to be removed from this list visit the *** *** CCP4 home page http://www.ccp4.ac.uk *** Hi Vince, you can try looking at the paper "Glycerol concentrations required for cryoprotection of 50 typical protein crystallization solutions" by Elspeth Garman and Ed Mitchell in J. Appl. Cryst. (1996). 29, 584-587 Amounts of glycerol are proposed to add to your PEG (8000) concentrations. Cheers, Virgile Quoting Vincenzo Carbone <[EMAIL PROTECTED]>: > Hi guys > > Sorry for the non ccp4 based question but i was wondering if anyone has any > tips on producing a suitable cryo buffer for crystals grown in peg8000. My > crystals come out quite quickly in 15 to 10% peg and can be quite large. Ive > tried to increase the peg concentration, replacing the peg with glycerol or > using a combination of peg8000 and glycerol but each time the crystals fail > to diffract when flash frozen. Previous experience with these crystals have > shown diffraction of up to 2A when dry mounted. > > thanks, > > > vince > Virgile Adam ([EMAIL PROTECTED]) European Synchrotron Radiation Facility Office 18.1.07 - Sector 18 6 rue Jules Horowitz, BP 220, 38043 Grenoble Cedex 9 France Tel: +33 (0)4 76 88 27 72 / Fax: +33 (0)4 76 88 26 24 ***There are 10 kinds of people: those who can count in binary and the others*** ------------------------------------------------- This mail sent through IMP: http://horde.org/imp/
