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Can IsoFast™ Bst Polymerase be used to blunt DNA, fill in 3′ overhangs or remove 5′ overhangs?
After a restriction enzyme reaction, the end parts of a DNA molecule could be left blunt, with a 5′ overhang (3′ recessed end) or a 3’ overhang (5’ recessed end).
IsoFast™ Bst Polymerase shows only 5’ to 3’ polymerase activity but no 5’ to 3’ or 3’ to 5’ exonuclease activity (i.e. the polymerase has no proofreading activity). Therefore, it can only be used to fill in 5’ overhangs (generating a blunt end), but not to remove 3′ overhangs or 5’ overhangs, as it lacks exonuclease activity.