converting overhang into blunt end - can i use Pfu instead Klenow? (Apr/24/2006 )

hi! i have problem in cloning... i have to convert my vector and insert into blunt end; because they do not compatible... the problem is my lab do not have Klenow but have Pfu polymerase. Did anybody know whether Pfu can generate blunt end? thx b4...

In theory you should be able to use the 5'-3' polymerase activity to fill-in a 3' overhang when dNTPs are there. (Same idea as T4) I just have never heard of anyone trying it with Pfu.

hi! i have problem in cloning... i have to convert my vector and insert into blunt end; because they do not compatible... the problem is my lab do not have Klenow but have Pfu polymerase. Did anybody know whether Pfu can generate blunt end? thx b4...

You also could use S1 Nuclease or Mung Bean Nuclease to get blunt ends...