Can I use intercalating dyes for real-time monitoring with TwistAmp®?

Yes. It is possible to use intercalating dyes to quantify and monitor the TwistAmp® reactions in progress. However, as with PCR, the dye typically binds to any double-stranded DNA, so primer noise can generate false positive signals. This noise is made worse as the dye is monitored at a low temperature – in PCR assays the temperature is often such that primer dimers are melted when intercalated dyes are measured. Moreover, the exonuclease present in the TwistAmp® exo kit will digest most of the amplification product during the reaction, and the use of intercalating dyes is not recommended with these kits.

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