Protocol for blunting ends by 3' overhang removal and fill-in of 3' recessed (5' overhang) ends using DNA Polymerase I, Large (Klenow) Fragment (M0210)
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- DNA should be dissolved in 1x NEBuffer or T4 DNA Ligase Reaction buffer supplemented with 33 μM each dNTP.
- Add 1 unit of Klenow per microgram DNA.
- Incubate for 15 minutes at 25°C.
- Stop reaction by adding EDTA to a final concentration of 10 mM and heating for 20 minutes at 75°C.