FAQs

Yes. Clara® Probe 1-Step Mixes (including Purple mixes and all passive reference dye formats) support fast cycling RT-qPCR, with successful amplification demonstrated using shortened cycling conditions.

For ultra-fast cycling on the Bio Molecular Systems Mic instrument, we have successfully tested reverse transcription at 52 °C for 2–5 minutes, followed by polymerase activation at 95 °C for 1-2 minutes and 40 cycles of 95 °C for 1–3 seconds and approximately 60 °C for 3–5 seconds. A pre-cycling step of 4 cycles of 95 °C for 1 second and 60 °C for 5 seconds can improve results by pre-amplifying the target.

Ultra-fast cycling requires instruments capable of rapid temperature changes and efficient fluorescence acquisition. The Bio Molecular Systems Mic is particularly well suited to ultra-fast cycling, while instruments such as the Roche LightCycler® 480 and fast-mode Applied Biosystems™ QuantStudio™ systems also support accelerated cycling protocols.

The optimal conditions depend on the target, amplicon length, reaction complexity and instrument. We recommend using short amplicons of no more than 120 bp and verifying amplification efficiency and sensitivity when shortening the reverse transcription or cycling steps, particularly for multiplex assays and low-copy RNA targets. For UNG-containing mixes, include the additional UNG incubation specified in the product manual.