HomeFAQsHow can I remove the remaining primers and dNTPs from a PCR reaction prior to DNA sequencing?
FAQ: How can I remove the remaining primers and dNTPs from a PCR reaction prior to DNA sequencing?
In a typical protocol, one can mix 5 µl of PCR reaction (from 30-35 PCR cycles), 1 µl of Thermolabile Exonuclease I and 1 µl Quick CIP (NEB #M0508, no need to add rCutSmart Buffer) together, followed by incubation at 37°C for 4 minutes, then at 80°C for 1 minute to inactivate the enzyme. Store the cleanup sample at -20°C prior sequencing. Use 3 µl of the cleanup product for Sanger-base sequencing.