Characterization of Quantitative FRET Sensors for Fluorescence Molecular Tomography

Qing Liu, Xin Xie, Yi Zhang, Yanyan Xu, Yue Zhang, Xin Liu, Jing Bai, Xiaodong Liu
2011 Biophysical Journal  
nanocluster formation, as these are essentially non-fluorescent. Our detection technique is simple, inexpensive, and compatible with commercial DNA synthesizers. It is also the first demonstration that a turn-on probe can be made based on fluorescent noble metal nanoclusters. Our initial investigation demonstrated NCB detection of an influenza target with a signal-to-background (S/B) ratio five times better than that of a conventional molecular beacon. Here, we expand upon this work to
more » ... te a method of using NCBs to differentiate single-nucleotide variations. Our method discriminates single-nucleotide variants by colorimetric change of the NCB probes rather than fluorescence intensity change. This added dimensionality enables the increase in fluorescence intensity (on/off switching) to quantify the amount of target, whereas the fluorescence color identifies single-nucleotide variants. Samples with single-nucleotide variations can be unambiguously identified on a common gel imager with naked eyes, making this method a reliable and low-cost assay with simple readout format.
doi:10.1016/j.bpj.2010.12.2847 fatcat:idq5dmx365c2tfwcm4db24haiy