A Typical Exonuclease V Reaction (M0345)
1. Setup the following reaction:
|COMPONENTS||50 μl REACTION|
|DNA||up to 1 μg|
|NEBuffer 4 (10X)||3 μl (1X)|
|ATP (10 mM)||3 μl (1 mM)|
|Exonuclease V||1 μl (10 units)|
|Nuclease-free H2O||to 30 μl|
2. Incubate at 37°C for 30 minutes.
3. To stop reaction add EDTA to 11 mM.
4. Heat Inactivation 70°C for 30 minutes.
5. Clean-up treated samples by column purification and/or ethanol precipitation.
Note: Estimate amount of DNA to be removed by agarose gel electrophoresis or OD260. If > 1 μg scale up all reaction components proportionately.