20× One Step qRT Enzyme Mix contains M-MLV (RNase H-) reverse transcriptase, RNase Inhibitor, Hot Start DNA Polymerase; 2× One Step qRT Buffer contains SYBR Green I, 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.