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When might I need to adjust the primer-to-probe ratio in my qPCR experiments?

In multiplex experiments, it may be useful to modulate the primer-to-probe ratios. While ratios that work best will depend on your particular experiments, a good starting point is to limit the primer-to-probe ratio for the highest expressing genes to 1:1 and increase the ratio for the lowest expressing genes to 4:1. Researchers may also want to adjust the ratio to reproduce experiments where specific primer:probe ratios have been published. IDT PrimeTime® qPCR Assays allow you to select a premixed primer/probe ratio from 1:1 to 4:1 to maximize experimental flexibility.