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