Protocol for T5 Exonuclease (M0363)

T5 Exonuclease will degrade ssDNA and linear and nicked dsDNA, leaving supercoiled dsDNA intact. Set-up the reaction as follows:

DNA x µl (1 µg)
10X NEBuffer 4 5 µl
T5 Exonuclease 1 µl (10 units)
H2O x µl (up to 50 µl)

  1. Incubate at 37°C for 30 minutes.
  2. To stop reaction, add EDTA to a concentration of 11 mM.
  3. Clean-up treated samples by column purification or phenol/chloroform extraction followed by ethanol precipitation.