Noise signal are very high when multiplex RT-PCR using 4 taqman probes FAM/ROX/HEX/CY5 ?

22 December 2015 1 6K Report

All components are freeze-dried. We use AB7500.

compare the liquid state, the other channels' noise signal of freeze-dried components are very high when we only detect one channel

Matthew Sobo

I would recommend testing the primers separately to make sure they are amplifying correctly. As you pointed out, it looks like only one channel is amplifying.

This one may seem obvious but... for your ROX line, make sure you are using a MM that doesn't include ROX as that would likely give you a high bit of background for that color.

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