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 and optimized buffer; 50× ROX Reference Dyes are used to rectify the error of fluorescence signals between different wells.
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.

Please select an appropriate ROX according to the Real-time PCR instrument used:
Do Not Use ROX:Bio-Rad CFX96™, CFX384™, iCycler iQ™, iQ™5, 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®.
Use ROX1 (1×):ABI 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast; StepOne™, StepOne Plus™.
Use ROX2 (1×):ABI 7500, 7500 Fast, Quant-Studio 3, 5, 6, 7, 12K Flex, Dx, ViiA™7; Stratagene MX3000P™, MX3005P™,MX4000™.