UltraPlex 1-Step ToughMix is a ready-to-use, 4X concentrated master mix 1-step reverse transcription and real-time quantitative PCR (RT-qPCR) of RNA templates using probe-based detection methods. First-strand cDNA synthesis and PCR amplification are carried out in the same tube without opening between steps. Optimized to deliver highly sensitive quantification of RNA viruses or low abundance RNA targets in uni- or highly multiplexed RNA detection assays, this reagent chemistry is optimized to deliver maximum assay sensitivity, precision and reproducibility with miniaturized reaction volumes and either conventional or accelerated thermal cycling protocols. UltraPlex 1-Step ToughMix contains all required components for RT-qPCR except RNA template and probe and is compatible with all dual-labeled probe chemistries.