The Reliance One-Step Multiplex Supermix is a highly sensitive and robust reagent mix for detection and quantification of up to five RNA or DNA targets in one reaction. This kit offers robust, sensitive amplification and detection of up to five targets under a wide range of conditions, including challenging templates, difficult targets, or targets that are in the presence of PCR inhibitors. With its room-temperature stability, the Supermix can be easily integrated into automated workflows. Reliance One-Step Multiplex Supermix is a ready-to-use, single-tube reagent mix optimized for sensitive amplification of up to five targets in a single reaction. It is formulated with an optimal enzyme blend, including a new Reliance reverse transcriptase and Sso7d fusion polymerase, and an advanced buffer to maximize sensitivity and efficiency in multiplex RT-qPCR reactions. Its extended assembled reaction stability (up to 24 hours) is adapted for high-throughput automated workflows.