RTGreen is a very sensitive dye for the detection of double stranded DNA (dsDNA). The dye is a green fluorescent nucleic acid dye with features that make it useful for non-specific detection of amplification in realtime qPCR experiments.
Compared with the widely used SYBR Green I, RTGreen dye is generally less inhibitory toward PCR and less likely to cause nonspecific amplification , RTGreen dye can be used at a much higher dye concentration than SYBR Green I, resulting in more robust PCR signal.
The PCR reaction can be monitored using our existing optical setting for SYBR Green I or FAM on any commercial real-time PCR cycler. The qPCR protocol provided below is for PCR using regular non-hot-start Taq. Use of a hot-start Taq may require some adjustment of PCR buffer composition in terms of ionic strength and pH to best take the advantage of RTGreen dye. The water soluble solvent such as DMSO or glycerol are frequently added to stabilize master mixes. These components and pH may need to be optimized depending on the enzyme used.