Protocol for T5 Exonuclease (M0363)
T5 Exonuclease will degrade ssDNA and linear and nicked dsDNA, leaving supercoiled dsDNA intact.
1. Set-up the reaction as follows:
|COMPONENTS||50 μl REACTION|
|DNA||up to 1 µg|
|NEBuffer 4 (10X)||5 µl (1X)|
|T5 Exonuclease||1 µl (30 units)|
|Nuclease-free H2O||up to 50 µl|
2. Incubate at 37°C for 30 minutes.
3. To stop reaction, add EDTA to a concentration of 11 mM.
4. Clean-up treated samples by column purification or phenol/chloroform extraction followed by ethanol precipitation.