Protocol Online logo
Top : Forum Archives: : Real-Time PCR

Using sybr green and Taqman probes on the same plate? - (Jul/16/2007 )

Hi there,

I would like to assay some genes using sybr green and also run the HKG with a taqman probe, preferably on the same plate.

The reason is that I have used both chemistries in the past for different projects, and have designed primer pairs that work quite well.
I would prefer to keep using the probe for my housekeeping gene, since I have developed it already, I know it is specific, and I still have stock.

The main issue I can see is that the program for sybr green normally requires a 72 degree extension step, instead of a combined 60 degree extension/annealing step for a probe.

Does anyone know whether it will cause a problem if I run the sybr green with a 60 degree combine extension/annealing step??



You can easiliy use SYBR green with a two step protocol. But at least our ABI Prism cycler does not allow to put SYBR and other detectors on the same plate.