2 replies to this topic

[Curtis]

I know that Taqman probes should not overlap with the primers when doing real-time pcr, but can you please tell me how far the probe and primers could be located from each other so that the PCR is efficient?

[TanyHark]

Curtis, on Feb 21 2009, 12:00 PM, said:

I know that Taqman probes should not overlap with the primers when doing real-time pcr, but can you please tell me how far the probe and primers could be located from each other so that the PCR is efficient?

I don't know but I generally keep at least 7-10 nt distance on both sides, and most of my primer-probe combos have worked.

[chrisbelle]

The primers should be designed as close as possible to the probe without overlapping it.