fillin - bluntend (Aug/16/2007 )
can I use T4 pol or klenow
to fillin a fragment cut with HindIII and Kpn1
i would rather use klenow
but with HindIII and Kpn1 they produce 5' overhang and 3' overhang.
klenow can fill 5' overhang and 3' overhang?
klenow will fill in 5' overhangs provided dNTP is available.
On the otherhand, klevow will remove 3' overhangs by 3' to 5' exonuclease activity.
Both will result in a blunt end.
For interest sake;
Asp718 is an isoshizozyme of Kpn. Both recognises the same site. However Asp718 produces 5' overhangs