Why is the initial fluorescence signal too high when using artus PCR assays on the Rotor-Gene cycler?

Manufacturing failure/faults or air bubbles in the tubes and/or capillaries may cause initial fluorescent signals that are too high. When using artus® kits designed for the Rotor-Gene instrument with a total volume of 50 μl, the 20 μl sample has to be mixed with 30 μl of artus® Master Mix in the reaction tube by pipetting up and down several times. Otherwise, distribution of fluorescence is not homogeneous within the first few PCR cycles which may lead to initial fluorescence signals that are too high.

 

 

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