Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

Removal of 5 overhangs to form blunt ends - (Jun/09/2010 )

Hi all,

Am having problems with a new construct and I need to put my protein of interest in frame, I need to remove the 5' overhangs generated when I digest with XhoI. Unfortunately, T4 polymerase exonuclease activity is 3'-5', I know Polymerase I has 5'-3' exonuclease activity but am confused, I don't know how to use it to cut these 5' overhangs, please HELP :wacko:



DNA polymerase I can not be used to remove 5'-overhangs ...use Mung bean nuclease instead!

Here you can find how to do it.