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Label-free single-molecule detection of DNA-hybridization kinetics with a carbon nanotube field-effect transistor

Abstract

Single-molecule measurements of biomolecules can provide information about the molecular interactions and kinetics that are hidden in ensemble measurements. However, there is a requirement for techniques with improved sensitivity and time resolution for use in exploring biomolecular systems with fast dynamics. Here, we report the detection of DNA hybridization at the single-molecule level using a carbon nanotube field-effect transistor. By covalently attaching a single-stranded probe DNA sequence to a point defect in a carbon nanotube, we are able to measure two-level fluctuations in the conductance of the nanotube in the presence of a complementary DNA target. The kinetics of the system are studied as a function of temperature, allowing the measurement of rate constants, melting curves and activation energies for different sequences and target concentrations. The kinetics demonstrate non-Arrhenius behaviour, in agreement with DNA hybridization experiments using fluorescence correlation spectroscopy. This technique is label-free and could be used to probe single-molecule dynamics at microsecond timescales.

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Figure 1: Electrochemical oxidation of a carbon nanotube.
Figure 2: Real-time measurements of DNA kinetics.
Figure 3: DNA melting curves.
Figure 4: DNA kinetics analysis.

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Acknowledgements

This work was supported in part by the National Science Foundation (grants ENG-0707748 and CHE-0641523). Additional support was provided by the New York State Office of Science, Technology, and Academic Research (NYSTAR). This work was also supported in part by the Office of Naval Research (grants N00014-09-01-0250 and N00014-09-1-1117) and by the National Institutes of Health (grant R33-HG003089).

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S.S., C.-Y.C. and K.L.S. designed the experiments. S.S. and C.-Y.C. performed the experiments and analysed the data. Y.-J.Y. and P.K. assisted in the AFM and SGM experiments. C.N and R.L.G. assisted with data analysis. S.S., C.-Y.C and K.L.S. co-wrote the paper. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Kenneth L. Shepard.

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The authors declare no competing financial interests.

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Sorgenfrei, S., Chiu, Cy., Gonzalez, R. et al. Label-free single-molecule detection of DNA-hybridization kinetics with a carbon nanotube field-effect transistor. Nature Nanotech 6, 126–132 (2011). https://doi.org/10.1038/nnano.2010.275

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