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I am currently redesigning some FRET probes and therefore I have the opportunity to optimize my FRET pair.

Due to limited lasers available I am restricted to 488nm (EGFP) as donor. So my question is which acceptor to use.

Does anyone know of direct comparisons between EGFP:mCherry and EGFP:TagRFP, and TagGFP:mCherry and TagGFP:TagRFP, and/or does anyone know a better FRET pair working with the 488nm/561nm lasers?

I am planning to compromise and do both sensitized FRET and fluorescent lifetime.

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2 Answers

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I found a FRET Pair Spectra. Hope it helps:

http://www.kcci.virginia.edu/FRET-FLIM/spectra/index.php

FRET Pair Spectra

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Maybe you could use organic dyes: Alexa488 & TMR (Couple via: snap-, halo-, Sfp-, Ap-tag).

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