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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
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