Currently I am trying to digest my plasmid using NEB SapI. There are 5 cutting sites in my plasmid. I found that the digestion is quite inefficient. Below is my reaction mixture:
Plasmid 1ug
SapI(10U/ug) 0.5ul
10xNEBuffer4 5ul
dH2O up to 50ul
SapI is quite expensive so I don't wish to use to much of them. Is there any suggestion for the protocol?
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