10.1021/bc2002659.s001 Andrea Kienzler Andrea Kienzler Roman Flehr Roman Flehr Rolf A. Kramer Rolf A. Kramer Sören Gehne Sören Gehne Michael U. Kumke Michael U. Kumke Willi Bannwarth Willi Bannwarth Novel Three-Color FRET Tool Box for Advanced Protein and DNA Analysis American Chemical Society 2011 peptide fluorescence probes building block approach life science applications DNA AnalysisWe report FRET interaction 2011-09-21 00:00:00 Journal contribution https://acs.figshare.com/articles/journal_contribution/Novel_Three_Color_FRET_Tool_Box_for_Advanced_Protein_and_DNA_Analysis/2612008 We report on a new three-color FRET system which we were able to verify in peptides as well as in synthetic DNA. All three chromophores could be introduced by a building block approach avoiding postsynthetic labeling. Additional features are robustness, matching spectroscopic properties, high-energy transfer, and sensitivity. The system was investigated in detail on a set of peptides as well as an array of tailored oligonucleotides. The detailed analysis of the experimental data and comparison with theoretical considerations were in excellent agreement. It is shown that in the case of polypeptides specific interaction with the fluorescence probes has to be considered. In contrast with DNA, the fluorescence probes did not show any indications of such interactions. The novel three-color FRET toolbox revealed the potential for applications studying fundamental processes of three interacting molecules in life science applications.