20× EZProbe One Step Enzyme Mix contains M-MLV (RNase H-) reverse transcriptase, RNase Inhibitor and Hot Start DNA Polymerase; 2× EZProbe One Step Buffer contains dNTPs, Oligo dT18, Random Hexamer, optimized buffer and ROX1.
The M-MLV Reverse Transcriptase, which is an RNA-dependent DNA polymerase with a significantly attenuated RNase H activity, can synthesize cDNA at a temperature range of 42°C to 60°C and is inactivated during the activation step of the Hot Start DNA Polymerase.
RNase inhibitor can protect RNA templates from degradation.
This buffer has been optimized to allow both reverse transcription and PCR amplification to occur in the same reaction across a wide range of templates.
Hot Start DNA Polymerase requires an activation step at 95°C for 5 minutes, which prevents non-specific amplification during cDNA synthesis.

Appropriate real-time PCR instruments for this kit are as follows:
Bio-Rad CFX96, CFX384,   iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon, Opticon 2,   Chromo4; Roche LightCycler 96, Roche   LightCycler 480; Eppendorf Mastercycler   ep realplex, realplex 2s; Illumina   Eco qPCR; Qiagen/Corbett   Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene   6000; Thermo Scientific PikoReal   Cycler; Analytikjena qTOWER 3G; Cepheid SmartCycler.
ABI 5700, 7000, 7300, 7700,   7900, 7900HT, 7900HT Fast; StepOne, StepOne Plus.