Fluorescence Resonance Energy Transfer (Fret) to Follow Ribozyme Reactions In Real Time

Andreas Hanne Institute for Hematopathology and Applied Cancer Research, Christian-Albrechts-University, Niemannsweg 11, D-24105, Kiel, Germany, & IntelliGene Ltd., Jerusalem, Israel.

Muktevi V. Ramanujam Institute for Hematopathology and Applied Cancer Research, Christian-Albrechts-University, Niemannsweg 11, D-24105, Kiel, Germany, & IntelliGene Ltd., Jerusalem, Israel.

Thomas RüUcker Institute for Hematopathology and Applied Cancer Research, Christian-Albrechts-University, Niemannsweg 11, D-24105, Kiel, Germany, & IntelliGene Ltd., Jerusalem, Israel.

Guido Krupp Institute for Hematopathology and Applied Cancer Research, Christian-Albrechts-University, Niemannsweg 11, D-24105, Kiel, Germany, & IntelliGene Ltd., Jerusalem, Israel.

Abstract

Fluorescence resonance transfer (FRET) was applied for real time monitoring of ribozyme reactions. Group I ribozyme ligation was followed with two separate, fluorescent-labeled RNA substrates. For hammerhead ribozyme cleavage, a double-fluorescent-labeled substrate was used. For the first time we analyzed multiple turnover conditions. Real time monitoring permits convenient analysis of ribozyme kinetics and the sequence-specific, quantitative detection of RNAs in femtomole amounts.

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Fluorescence Resonance Energy Transfer (Fret) to Follow Ribozyme Reactions In Real Time

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Fluorescence Resonance Energy Transfer (Fret) to Follow Ribozyme Reactions In Real Time

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