No Template Control Contamination qReal Time PCR ?

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No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

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Cristian Ricaurte

Yes I tried. I am running 40 Cycles. CT value around 24 - 28. Will you suggest start from scratch or change primer concentration ?

Ali Akgul

I think you better test your negative control with regular PCR see how it looks like. Also reduction of primer concentration would help. You can test both in PCR before even run a qrt Pcr plate.

Best luck in your experiments...

Justyna Augustyniak

Hi,

The melting curve after qPCR cycle can show a product in NTC in the situation when you get homodimer and/or heterodimer formation which are a result of using incorrect design primers in qPCR reactions.

The homodimer and/or heterodimer formation are also generated when the primers concentration is too high.

1) Guidelines for Successful Quantitative Gene Expression in Real- Time qPCR Assays, https://www.intechopen.com/books/polymerase-chain-reaction-for-biomedical-applications/guidelines-for-successful-quantitative-gene-expression-in-real-time-qpcr-assays

Good luck!