Judging a dissociation curve - (Jun/15/2009 )
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.
I'd be a little concerned about H4. Was it a control reaction or a sample?
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?