Yes. Some optimization may be required in your system.
The CleanAmp™ One-step RT-PCR Master Mix provides a Hot Start RT step, which prevents amplification of unwanted side reactions and truncated products due to RNA secondary structure. At 47°C, a small amount of denatured RNA template will be present to generate just enough of the correct cDNA to go into the PCR reaction. A key step in developing the CleanAmp™ One-step RT-PCR Master Mix was ensuring amplification of targets with high secondary structure, such as ABCA7 and PBGD.
The RT enzyme has an optimal temperature range of 42°C – 48°C. Due to the CleanAmp™ deprotection kinetics, we recommend temperatures at the higher end of that range, optimally 47°C.
Yes. We have successfully amplified up to 4 targets in the same reaction. Higher plexing may be possible.
Yes. In addition to primers, template RNA and water, add real-time reagents according to manufacturer’s recommendations (SYBR® Green, SYTO® 9, TaqMan® probes, ROX™, etc.)