How do you localize qPCR inhibition before redesigning primers?
by Lale
When amplification efficiency is acceptable on clean template but poor in the intended sample matrix, what experiment do you run next to locate the inhibition? Please give the order in which you test sample dilution, template cleanup, an exogenous amplification control, and a matrix spiked with a known target. Which pattern implicates the matrix strongly enough to retain the primer pair, and which result still sends you back to primer redesign?
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