Premix Ex Taq (Probe qPCR) is a 2X master mix for real-time PCR (qPCR) using probe-based qPCR or 5' nuclease assays. This 2X master mix includes Takara Ex Taq HS—a hot-start PCR enzyme in combination with anti-Taq antibody—in a qPCR-optimized buffer. Takara Ex Taq HS inhibits non-specific amplification while enabling high-efficiency amplification and detection sensitivity during real-time PCR analyses. Additionally, Tli RNase H, a heat-resistant RNase enzyme, is included in the real-time PCR premix in order to minimize PCR inhibition due to the presence of residual mRNA in the input cDNA. This master mix is ideal for high-speed PCR, allows accurate target quantification and detection over a broad dynamic range, and enables highly reproducible and reliable real-time PCR analyses.