I echo this. Some time ago I was working on a target which seemed to precipitate when cleaving overnight with TEV. I wasted a fair bit of time trying to optimise cleavage conditions with a myriad of buffers. In the end, just by not agitating my solution, there was no precipitation and recovery was extremely good.
I never agitate my solutions when cleaving now and have not had any problems with cleavage (to do with this issue, anyway) or recovery. cheers charlie Dr. Charles Allerston Genome Integrity Group Structural Genomics Consortium Nuffield Department of Medicine Old Road Campus University of Oxford OX3 7DQ http://www.sgc.ox.ac.uk/ -----Original Message----- From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Xiaopeng Hu Sent: 31 March 2011 15:23 To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Question about TEV cleavage Our experience is do not shake the tube during TEV cleavage,I dont know why, but it does help. xiaopeng
