I can’t see to find in the literature how a gel electrophoresis or melting curve analysis of ddPCR would be possible. Assuming it is not, I am wondering how can unspecific amplifications then be detected? I would appreciate any helpful thought.
Hi,
My first step is generally to check the primers and probe sequences on an nucleotide alignment to make sure they only match the right target, and my second step is to test the assay on extracts of closely-related targets, which should not lead to any amplification...
But I guess you could push it further if you are really worried about your samples, and you could technically run a normal PCR with your primers and probes and do a gel electrophoresis? I'm guessing you could even sequence the PCR products if you want to know exactly what is in there?
The last bit is theoretical but hopefully something will help solve your issue!
Kind regards, Maïlys
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