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.