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Nucleic Acids Res. 2014 Apr;42(8):e65. doi: 10.1093/nar/gku111. Epub 2014 Feb 7.

Thioflavin T as a fluorescence light-up probe for G4 formation.

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ARNA Laboratory, University of Bordeaux, F-33000 Bordeaux, France, INSERM U869, IECB, F-33600 Pessac, France and Department of Chemistry and Biochemistry, Swarthmore College, Swarthmore, PA 19081, USA.


Thioflavin T (ThT) becomes fluorescent in the presence of the G-quadruplex structure such as that formed by the human telomeric motif. In this report, we extend and generalize these observations and show that this dye may be used as a convenient and specific quadruplex probe. In the presence of most, but not all, G4-forming sequences, we observed a large increase in ThT fluorescence emission, whereas the presence of control duplexes and single strands had a more limited effect on emission. This differential behavior allowed us to design a high-throughput assay to detect G4 formation. Hundreds of different oligonucleotides may be tested in parallel for G4 formation with a simple fluorescence plate reader. We applied this technique to a family of aptamers not previously recognized as G4-forming sequences and demonstrated that ThT fluorescence signal may be used to predict G4 formation.

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