Protocol for Nuclease BAL-31

Nuclease BAL-31 is used for progressive shortening of linear duplex DNA.*

 1. Set-up the reaction as follows:

COMPONENTS For 50 μl Reaction
 DNA Approximately 3 picomoles of ends
 Nuclease BAL-31 Reaction Buffer (2X) 25 μl (1X)
 BAL-31 Nuclease 1 μl (1 unit)
 Nuclease-free H2O  up to 50 μl

2. Incubate at 30°C for 10 minutes.

3. Stop reaction by adding EGTA to at least 30 mM. 

4. Heat Inactivation 65°C for 10 minutes.

5. To cleanup treated samples we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit, NEB #T1030), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit, NEB #T1020), or performing a phenol/chloroform extraction followed by ethanol precipitation. 

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