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.