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Respected All, Thanks for your valuable suggestions and inputs. with regards, Harsh On Thu, Mar 21, 2013 at 7:38 AM, Bosch, Juergen <[log in to unmask]> wrote: > 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 > > > > >