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.