qPCR - no amplification curve but suitable melting curve - (Jul/18/2013 )
Hi there,
last week i run a qpcr-probe (two step, 40 cycles); there was no amplification , then i analysed the melting curve an there was a really suitable peak!
what´s going wrong there? I would be really grateful to have suggestions from you!
greets,
Martin
-pcr_martin-
pcr_martin on Thu Jul 18 07:59:52 2013 said:
Hi there,
last week i run a qpcr-probe (two step, 40 cycles); there was no amplification , then i analysed the melting curve an there was a really suitable peak!
what´s going wrong there? I would be really grateful to have suggestions from you!
greets,
Martin
Do you still have the plate? You could put the reaction on to an agarose gel and see if there is an amplicon as indicated by the melting curve.
-Podge-
Hi,
I´ve done it, there was a amplicon as indicated; attached you´ll find a scrrenshot; the red peak is the first band, the green one the second band
-pcr_martin-
It seems as though your probe is not binding.
-biothreat-