Volcano3G RT-PCR Probe 2x Master Mix works very well also for DNA amplification assays. This master mix is optimized for an amplicon size between 60- 300 bp.Minimize the number of freeze-thaw cycles by storing in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4°C.
- Run a PCR with a temperature gradient at the RT-step and annealing step I in order to find the optimal temperature for your assay.
- Most RT-PCR assays work well with a RT-cycling step consisting of a short denaturation followed by incubation at 58-70°C and subsequent PCR cycling.
- Rapid detection and identification of RNA & DNA targets
- Reverse transcription qPCRs (RT-qPCRs)
Quality Control Assays:
RT-PCR activity: Volcano3G RT-PCR Probe Mix is tested for a successful RT-qPCR performance. A 151 bp fragment (HPRT1 mRNA) is amplified from human total RNA extract and the linearity of amplification over a specified serial dilution is demonstrated. The activity of the Volcano3G DNA polymerase is monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and DNA primer. Enzyme concentration is determined by protein-specific staining. Please inquire more information at [email protected] for the lot-specific concentration.
No contamination has been detected in standard test reactions.
Volcano3G is the next generation of our Volcano2G with an improved performance. It offers a variety of applications for a precise and rapid qPCR. It is applicable with common thermal cycling protocols and “zero-step” RT-PCR protocols direct from RNA. As demonstrated here, artificial HPRT1 mRNA was detected in a 10-fold dilution series by using a probe-based qPCR assay and a common thermal RT-PCR protocol.
Be quick! No isothermal reverse transcription step is needed: A fast start function due to a hotstart aptamer and the engineered, truly thermostable Taq DNA polymerase with reverse transcriptase activity allows immediate cycling.
Even directRT-PCRs are possible like this: Sample extraction and sample lysis is not necessary as immediate hot PCR cycling conditions can brake cell- and virus membranes. Please see our published references to this product