Hi Jacob, Keep regular drops and run a cat whisker through the drops after 4 - 5 hours. You should have small crystals within 6-10 hrs. If they are still large, use higher protein concentration or precipitant.
Kind regards Mathews -----Original Message----- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Jacob Keller Sent: Thursday, July 03, 2008 4:08 PM To: [email protected] Subject: [ccp4bb] Small lysozyme crystals protocol Dear Crystallogrphers, does anybody here know a protocol to get consistently well-diffracting but smaller, ~50um, cryoprotect(ed/able) lysozyme crystals? Thanks, Jacob Keller ******************************************* Jacob Pearson Keller Northwestern University Medical Scientist Training Program Dallos Laboratory F. Searle 1-240 2240 Campus Drive Evanston IL 60208 lab: 847.491.2438 cel: 773.608.9185 email: [EMAIL PROTECTED] ******************************************* ----- Original Message ----- From: "Ailong Ke" <[EMAIL PROTECTED]> To: <[email protected]> Sent: Thursday, July 03, 2008 4:13 PM Subject: [ccp4bb] air bubbles in the Bio-Rad Duoflow system > Hello All, > > I own a Bio-Rad Duoflow system for almost a year. The machine lived up to > some of the recommendations I saw on this board. However, there is this > annoying problem with air bubbles entering columns and I cannot figure out > the source. The system would be free of any air bubbles in the beginning, > and performs fine when water is loaded using needle/syringe into the 5ml > superloop and then injected into the column. When protein samples are > injected the same way, a lot of air bubbles would appear and get trapped > inside the ion exchange column, leaving yellowish marks on the Uno columns > and eventually reducing their performance. I've been a FLC/AKTA user for > about ten years and have never seen such problems. I doubt it is due to a > faulty injection valve because we recently had it replaced for other > reasons. We did move a backpressure generator (a little black piece) from > post-column position to before-column, otherwise we cannot run sizing > columns in a reasonable flow rate. > > I'd like to hear if you have similar experiences and how you fixed the > problem. Thanks a lot! > > Ailong > > -- >
