site after blunting - (Jun/21/2007 )
can anybody please teach me how you calculate the constructed site after blunting of vector and insert digested with a different enzyme
for example if i digest my vector with speI and my insert with speI and kpnI then after blunting and ligation of the insert in vector what site will be construct in the both ligated site that is blunted speI-speI and blunted speI-kpnI??
When you blunt ends, you may add an extra base or remove extra base. So usually a matter of a single base. Best would be to sequence the plasmid once you have it to verify the exact sequences.