FIG. 1.

Molecular beacon strategy for Oct4 detection. (A) Schematic model for dual fluorescence resonance energy transfer (FRET) molecular beacon system. 6FAM and 5ROX fluorophores were chosen for their spectral overlap (Ex 490 nm/Em 520 nm and Ex 580 nm/Em 610 nm, respectively). BHQ1 and BHQ2 were used to quench 6FAM and 5ROX emissions, respectively, in solution. (B) Prediction of Oct4 mRNA secondary structure using RNAfold (http://rna.tbi.univie.ac.at/cgi-bin/RNAfold.cgi). Gray circle highlights region amenable to molecular beacon design. (C) Absorption/emission spectra for 6FAM and 5ROX demonstrating overlap between 6FAM emission and 5ROX absorption spectra with >30% response at 540–560 nm. Color images available online at www.liebertonline.com/scd.