Why is there no difference in Ct value for serial dilutions of template mutant DNA sample done in constant concetrations of wildtype DNA template?

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Jixing Luo

This is a standard curve of G12A. 300nM refers to the blocker concentration. 20000 G12A represents the G12A template concentration added in the mastermix which is 20k per microlitre.

Katie A Burnette

Looks like your assay isn't optimized. I'm guessing your primer efficiency isn't between 95-105%. It's theoretically possible, but often not practical to detect less than 10 copies of a DNA template in a PCR reaction. Go back and optimize your primers.

Thank you for your answer. I will go and check that immediately. Cheers