Klenow Fragment, exo-, is the large fragment of DNA polymerase I . It exhibits 5'→3' polymerase activity, but lacks the 3'→5' and 5'→3' exonuclease activities of DNA Polymerase I. The 3'→5' exonuclease activity of the enzyme is eliminated by mutations in the 3'→5'-exonuclease active sit.
- Lacks 3'→5' exonuclease activity
- Incorporates modified nucleotides (e.g., Cy3-, Cy5-, fluorescein-, rhodamine-, aminoallyl-, biotin-labeled nucleotides)
- Active in restriction enzyme, PCR, and RT buffers
|metal chelators, PPi, Pi (at high concentrations) (7)|
|E.coli cells with a cloned DNA fragment of the mutated polA gene|