Has anyone tried to use the same quencher for 2 reporter dyes in multiplex qPCR?

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No title

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Hector D De Paz

It is possible, of course, but I recommend you to use other quencher such as BBQ or BHQ. TAMRA, as a quencher, is the worst option.

Regarding the fluorophore maybe other options (FAM and VIC, o FAM and Cy5) could be possible as well.

Good luck!

Ayodeji Stephen Owati

Thanks Hector for the answer. However, the filter in 7500/7500 fast system is not compatible with some of the quenchers (BBQ or BHQ). In addition, which platform do you use for probe synthesis, also recommend the quencher for Cy5

M N Manoj

You can use BHQ quenchers in ABI 7500 systems. It is not required to select the quencher - you can select the option "none". It works. As Hector mentioned BHQ series quenchers are much better than TAMRA. Also check for spectral overlap between the excitation/emission of fluorophores, when you are multiplexing.