HomeFAQsI've digested my DNA and now want to blunt directly without purifying, can I add the blunting reagents directly to the restriction digest?
FAQ: I've digested my DNA and now want to blunt directly without purifying, can I add the blunting reagents directly to the restriction digest?
We recommend to heat inactivate the restriction enzyme after a restriction digest and prior to the blunting reaction. If the restriction digest has been carried out with enzymes that can be inactivated by a heat treatment, then after the heat inactivation step, add 0.1 volume of dNTPs and 1 ul of Blunting Enzyme and incubate at room temperature for 15 minutes. If the restriction enzymes used are not inactivated by a heat treatment, then the reaction will need to be purified using a commercial purification kit (we recommend the Monarch PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.