Yes. Luna qPCR can be set up at room temperature, but for best results, reactions should be kept on ice prior to thermocycling. The DNA polymerase present in the qPCR Master Mix, NEB’s Hot Start Taq DNA Polymerase (NEB #M0495), contains a unique modified aptamer (SOMAmer^®) for temperature-dependent activity control, and is inhibited below 45°C. This reversible inhibition prevents undesired nonspecific activity until the reaction is heated above the release temperature during the initial denaturation step of PCR cycling.