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:

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.