Newbie Questions

p_u_c_k_o at p_u_c_k_o at
Mon Jun 6 10:55:58 EST 2005

Hey ppl, got some newbie question (roll eyes if u gotta):
For a double digest, when the optimal pHs of the two restriction
enzymes are different, do you do the higher pH (less acidic) reaction
first, then add a little acid to meet the second optimal pH.  Does
Tris-HCl play any part in the acidity of the buffer?
Also, I've heard you can exchange NaCl with a mixture of NaCl/KCl (as
long as the total concentrations remain the same), can you replace the
NaCl completely with KCl?

These are probably really dumb questions, I don't work in a lab or do
double-digests in real life, just doing a microbiology question sheet.


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