Protocol for Lambda Exonuclease (M0262)

Lambda Exonuclease efficiently degrades 5' phosphorylated linear dsDNA from 5' to 3' direction, leaving supercoiled dsDNA intact.*

1. Set-up the reaction as follows:

Components 50 μl REACTION
 DNA  up to 5 μg
 Lambda Exonuclease Reaction Buffer (10X)
 5 μl (1X)
 Lambda Exonuclease
 1 μl (5 units)
 Nuclease-free H2O
 up to 50 μl

2. Incubate at 37°C for 30 minutes.

3. Stop reaction by adding EDTA to 10 mM.

4. Heat Inactivation 75°C for 10 minutes.

5. To clean up treated samples, we recommend using one of the following steps:

a. Column clean up (we recommend the Monarch® PCR & DNA Cleanup Kit, NEB #T1030) or

b. Running the reaction on an agarose gel, and then extracting the DNA (we recommend the Monarch Gel Extraction Kit, NEB #T1020), or

c. Performing a phenol/chloroform extraction followed by ethanol precipitation.

*Note: For more precise results we recommend titration of the enzyme to the intended substrate.