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Strange Taqman probe negative fluorescence

real-time PCR taqman fluorescence problem

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2 replies to this topic

#1 Rodriguez Ruslan

Rodriguez Ruslan


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Posted 01 April 2016 - 05:50 AM

Dear colleagues,

I'm observing strange behavior of my Taqman probes during the very first cycles of PCR: fluorescence becomes negative first, on later cycles things going more usual, but that first pitch spoils the whole picture. (see picture)

I’ve tried to find out what is that but failed, anyone knows anything?

Thank you a lot in advance!


Edited by Rodriguez Ruslan, 01 April 2016 - 05:51 AM.

#2 Andrea Fortina

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Posted 21 June 2016 - 08:09 PM

I don't why this happened. When I did qPCR, it seems had similar picture, but had no influence for the last result.

Andrea Fortina

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CreativeBiomart, Recombinant protein expert



#3 Trof


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Posted 26 June 2016 - 10:33 AM

The raw signal probably undergoes some technical normalization before showning in the graph, it obviously has no effect on the amplification plot so I would not worry about it. such early cycles are not even used for background normalization (or this may be actually also after background normalization, which takes zeroes the pre-amplification line).

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Also tagged with one or more of these keywords: real-time PCR, taqman, fluorescence, problem

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