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Judging a dissociation curve


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#1 Pluie

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Posted 15 June 2009 - 09:25 AM

Dear All,


Attached is a dissociation curve of a real-time PCR ran on a stratagene machine MX3000. Does it look alright, would you trust results from such a curve? The sequence amplified is a Beta-actin sequence.

As you can see only the signal H4 has a secondary peak higher than the product peak.


Dissociation_Curve3_BetaActin.jpg

Thanks,

P.

#2 gfischer

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Posted 15 June 2009 - 10:19 AM

I'd be a little concerned about H4. Was it a control reaction or a sample?
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#3 Pluie

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Posted 15 June 2009 - 10:27 AM

H4 is a sample. Negative samples were negative. The four highest peaks corresponds to the genomic control at 4 different concentrations.

If we ignore H4, what about the rest of the curve, is it acceptable to use the Ct values?

Edited by Pluie, 15 June 2009 - 10:32 AM.





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