FAQ: Can I use longer targets in one-step RT-qPCR?

In general, smaller amplicons (70 bp-200 bp) tend to result in higher efficiencies and earlier Cqs. For targets in this size range, a 30 second extension time (60 seconds on ABI machines) is recommended. Longer targets will benefit from longer extension times but the actual time will depend on the length of the target and instrument type, and should be determined empirically.