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Using primers meant for DIG probe as PCR diagnostic - (Jun/08/2006 )

I also posted this in General Techniques before I realized my mistake...

I was wondering if there are different criteria for the primers that are used in making DIG in situ hybridization probes vs. primers used in regular PCR?

In some of the papers I read, it seems that the same group will publish a different set of primers for in situ hybridization than they do for their regular PCR, but sometimes they use the same primer set for both purposes. My guess is that if it is Real Time PCR the primers might have to produce a smaller amplicon, but if I'm just performing regular PCR, and I'm making the probe using PCR anyway, why not use it as a standard primer set for PCR diagnostics.

Please let me know what you think about this.


i think if the set meets your requirements for both amplicons (dig probe and normal pcr) - eg size etc then it is fine......not sure about real time though