Encoding Multiple Virtual Signals in DNA Barcodes with Single-Molecule FRET

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DNA barcoding provides a way to label a myriad of different biological molecules using the extreme programmability in DNA sequence synthesis. Fluorescence imaging is presumably the most easy-to-access method for DNA barcoding, yet large spectral overlaps between fluorescence dyes severely limit the numbers of barcodes that can be detected simultaneously. We here demonstrate the use of single-molecule fluorescence resonance energy transfer (FRET) to encode virtual signals in DNA barcodes using conventional two-color fluorescence microscopy. By optimizing imaging and biochemistry conditions for weak DNA hybridization events, we markedly enhanced accuracy in our determination of the single-molecule FRET efficiency exhibited by each binding event between DNA barcode sequences. This allowed us to unambiguously differentiate six DNA barcodes encoding different FRET values without involving any probe sequence exchanges. Our method can be directly incorporated with previous DNA barcode techniques, and may thus be widely adopted to expand the signal space of DNA barcoding.
Publisher
AMER CHEMICAL SOC
Issue Date
2021-02
Language
English
Article Type
Article
Citation

NANO LETTERS, v.21, no.4, pp.1694 - 1701

ISSN
1530-6984
DOI
10.1021/acs.nanolett.0c04502
URI
http://hdl.handle.net/10203/281722
Appears in Collection
PH-Journal Papers(저널논문)
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