Yep, mostly you should stay away from Tris as this is the worst buffer system when playing with temperature changes. Tris for example has a ∆pKa/10˚C -0.31
Good, N.E. (1986) Biochemistry 5, 467 Jürgen P.S. @Matthew, was this what you meant by "the Good buffers often not" ? or just a coincidence ? On Mar 20, 2013, at 9:57 PM, Matthew Merski wrote: One of the other things you need to be concerned about with thermal melts is the change in buffer pKa as temperature varies (I seem to remember this being called the "beta" factor). Phosphate is used for CD melts regularly because its pKa is fairly invariant with temperature. (A good reference is "Data for Biochemical Research" by Dawson, Ch. 18). Acetate also shares this invariance but the Good buffers often do not. This is of course a concern with the Spyro Orange experiment as well. Matthew Merski Shoichet Group UCSF ...................... Jürgen Bosch Johns Hopkins University Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Office: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-2926 http://lupo.jhsph.edu
