Probes

[carobiotech]

Hi! does anyone know if I can design a probe using HEX and the quencher TAMRA?? I mean, usually I design mi probes with VIC and TAMRA and I wanted to know if I can change VIC for HEX and the result is the same. thanks!

[rowczarzy]

It depends on your equipment and the application. Is it for real-time PCR? Both VIC and HEX emits at 550-555nm, so the fluorescence emission and the TAMRA quenching is similar. It should be possible to change VIC to HEX. You may need to run a calibration for your PCR system to properly adjust for the HEX dye.

Is there a reason to use TAMRA for quenching? You may be better off to use dark quenchers (e.g. Iowa Black FQ). Because TAMRA emit fluorescence, your signal to background ratio is lower with TAMRA than with dark quenchers.

See fluorescent dye guide at
biophysics.idtdna.com/pdf/Fluorescence_and_Fluorescence_Applications.pdf

Feel free to email to the IDT customer service if you have more questions ([Only registered users see links. ]). They will help you.

Richard