enzyme that removes 5' overhangs - (Jun/20/2006 )
i am trying to make a mutant of a gene and i have to do a double digestion with bsiWI (c'gtac) , in the insert and hind III (a'agctt) in the vector, remove the intervening region and join back the two ends. i can use an adaptor or do blunt ligation. the problem isif i fill in the 5' overhangs created by these two enzymes, the myc tag is not in frame . i m using the pcdna3.1A(-)-myc/his tag vector from invitrogen.
i would be glad if anyne could suggest me a strategy without the use of an adaptor .
I think Mung Bean Nuclease is what you might be looking for. Check the Link.