Crush, pippet up and down, dilute as needed (I used 1:200) and vortex. Actually anything works... as everything - protein dependent.
For the stabilizing solution I have used the same as the reservoir solution (20%MPD, 250 mM NaCl, 0.1 M HEPES pH 7.5) or 2% more MPD. Worked fine and the seeds were usable more than 2 months (although I preferred freshly prepared). Dip a hair in the seeds and streak with it through three (or more) prepared drops in a row in order to obtain a dilution series of the seeding. So you can observe if the number of seeds makes any difference. Don't forget to test also variations on your conditions in the drops to be seeded (precipitant, pH, protein concentration). This might bring some surprising results! Good luck, Djordje >> Dear CCP4bbers, >> >> Please comment on the stabilizing solution for seed stocks. If the >> crystal >> is in 30% MPD and coming after 2-3 days, what should be the stabilizing >> solution for seeding. >> Thanks. >> James >> > > > -- > Dr Charles Allerston > Associate Research Scientist > Structural Biology > Ark Therapeutics > Darwin Building, UCL > Gower Street, > London WC1H 9EH > United Kingdom > > mailto: c.allers...@ucl.ac.uk > phone: +44 (0)2076791354 > Web: www.arktherapeutics.com > >
