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fillin - bluntend (Aug/16/2007 )

HI
can I use T4 pol or klenow
to fillin a fragment cut with HindIII and Kpn1

tanks

-ulujm-

i would rather use klenow

-fred_33-

klenow.

-perneseblue-

ok
but with HindIII and Kpn1 they produce 5' overhang and 3' overhang.
klenow can fill 5' overhang and 3' overhang?

-ulujm-

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

-perneseblue-