A Typical Exonuclease V Reaction (M0345)

Introduction

x μl sample DNA (~ 1 μg)
3 μl NEBuffer4 (10X)
3 μl 10 mM ATP
y μl H20 (up to final volume of 30 μl)
1 μl Exonuclease V (10 units)

Protocol

  1. Incubate at 37°C for 30 minutes.
  2. To stop reaction add EDTA to 11 mM.
  3. Heat Inactivation 70°C for 30 minutes.
  4. 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.