한빛사 논문
Sung Hyun Kim§†‡, Hyunwoo Kim∥‡, Hawoong Jeong∥ Tae-Young Yoon§*
§School of Biological Sciences and Institute for Molecular Biology and Genetics, Seoul National University, Seoul 08826, South Korea
∥Department of Physics, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, South Korea
†(Present address) Department of Bionanoscience, Kavli Institute of Technology, Delft University of Technology, 2629HZ, the Netherlands
‡These authors contributed equally.
*Corresponding author
Abstract
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.
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