Role of restriction and TAE Buffer - (Oct/10/2005 )
May I know the components and their roles of restrction buffer and TAE buffer.
Thanks in advance.
i've poicked for example the NEB buffers
Buffer Compositions (1X):
10 mM Bis Tris Propane-HCl, 10 mM MgCl2, 1 mM DTT (pH 7.0 at 25°C).
NEBuffer 2 (blue):
10 mM Tris-HCl, 10 mM MgCl2, 50 mM NaCl, 1 mM DTT (pH 7.9 at 25°C).
NEBuffer 3 (red):
50 mM Tris-HCl, 10 mM MgCl2, 100 mM NaCl, 1 mM DTT (pH 7.9 at 25°C).
NEBuffer 4 (green):
20 mM Tris-acetate, 10 mM magnesium acetate, 50 mM potassium acetate, 1 mM DTT (pH 7.9 at 25°C).
Tris is a buffering system which pH is adjusted by HCl or acetate.
salts are required for enzymatic reaction
DTT is used for preservation of the restriction enzyme (not sure about that point but as DTT reuces the disulfure bonds... it should prevent activity of other proteins excepted restriction endonuclease...)
TAE : tris acetate EDTA
tris is buffer
acetate for pH adjustment
EDTA chelates divalents cations.
But google would have helped you shorter !