The Luna Probe One-Step RT-qPCR Mix with UDG (No ROX) is supplied at a 4X concentration and enables higher amounts of sample input which is relevant for applications where RNA present in low abundance is of interest such as pathogen detection. The mix consolidates the components for one-step RT-qPCR into a single tube including Luna WarmStart RT Hot Start Taq DNA Polymerase dNTPs and Murine RNase Inhibitor in an optimized buffer. Combining Hot Start Taq DNA Polymerase with a novel WarmStart-activated reverse transcriptase allows for dual control of enzyme activity via reversible aptamer-based inhibition. This temperature-dependent activation helps to prevent undesirable non-specific priming and extension prior to thermocycling providing added security for setting up reactions at room temperature. This formulation contains no reference dye and is compatible with any instrument that does not require ROX (if ROX normalization is needed ROX can be added.)