Providing Klenow fragment (DNA polymerase I Large Fragment)

Klenow fragment (DNA polymerase I Large Fragment) is the Large Fragment of DNA Polymerase I, E.coli. 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 site.

Add 10 mM EDTA to chelate the Mg2+ cofactor, which protects the DNA ends as they "breathe" while the temperature is increased. Then heat at 75°C for 20 minutes.

Klenow fragment (DNA polymerase I Large Fragment) is nice gene product.

Application ares:

1. Random-primed DNA labeling, see Protocol for Random-primed DNA Labeling.

2. Labeling of double-stranded DNA recessed 3'-termini, see Protocol for Labeling Recessed 3'-termini of Double-stranded DNA.

3. Strand displacement amplification (SDA).

4. DNA sequencing by the Sanger method.

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