I want to develop an assay about virus studies. So I am searching a method to detect simultaneously Venizella zooster, Japanese encephalitis, Cytomegalo virus and Epstein barr virus using only single PCR assay and gel electrophoresis.
Hi,
the only method I can think of that matches your demands is multiplex qPCR. You have to make sure that your amplicons all have different sizes if you want to subsequently seperate them on a gel.
Or you can try multiple primer pairs in one regular PCR setup with again different sized amplicons. I any way it requires a lot of testing and tweaking. If you want to use this method regularly as a lab staple and your sample is scarce it might be worth it. Otherwise I would recommend go singleplex.
https://lifescience.roche.com/documents/Step-by-Step-Protocol-for-Multiplex-PCR-with-the-FastStartHigh-Fidelity-PCR-System-and-the-PCR-Optimization-Kit.pdf
Best regards
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