Protocol Online logo
Top : New Forum Archives (2009-): : PCR, RT-PCR and Real-Time PCR

distance between probe and primers in Taqman method - how far should they be? (Feb/21/2009 )

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?

-Curtis-

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.

-TanyHark-

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

-chrisbelle-