Our PCR mixes work reliably with 10 or more copies of template per reaction but not lower than that. Keep the extracted DNA in the lowest volume possible and don’t dilute further after inactivating the protease. We advise setting up the assay with more cells and once the conditions are established, scaling it down from there.

Yes, it can be used to amplify DNA from bacterial colonies or bacterial cell lysate however, it’s not necessary to use the Rapid Extract PCR Kit in order to perform routine colony PCR. If you’re working from bacterial colonies use a sterile tip to pick a colony and resuspend into the 50µl PCR reaction. If working from liquid culture add 5µl of overnight culture to the final mix. Follow the general protocol and increase the initial denaturation time to 10 min at 95°C. For qPCR, it may be important to extract DNA with the PCRBIO Rapid Extract Lysis Kit to decrease background noise and control the amount of DNA added to the reaction.