Abstract
The spatial control of DNA and of self-assembled DNA constructs is a prerequisite for the preparation of DNA-based nanostructures and microstructures and a useful tool for studies on single DNA molecules. Here we describe a protocol for the accumulation of dissolved λ-DNA molecules between planar microelectrodes by the action of inhomogeneous radiofrequency electric fields. The resulting AC electrokinetic forces stretch the DNA molecules and align them parallel to the electric field. The electrode preparation from off-the-shelf electronic components is explained, and a detailed description of the electronic setup is given. The experimental procedure is controlled in real-time by fluorescence microscopy .
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Carbone A, Seeman NC (2002) Circuits and programmable self-assembling DNA structures. Proc Natl Acad Sci U S A 99:12577–12582. https://doi.org/10.1073/pnas.202418299
Rothemund PWK (2006) Folding DNA to create nanoscale shapes and patterns. Nature 440:297–302. https://doi.org/10.1038/nature04586
Niemeyer CM (2007) Functional devices from DNA and proteins. Nano Today 2:42–52. https://doi.org/10.1016/S1748-0132(07)70058-0
Bier FF, Hölzel R (2008) Nucleic acid based nanostructures—recent advancements and the impact of NUCAN. AIP Conf Proc 1062:3–12. https://doi.org/10.1063/1.3012301
Washizu M, Kurosawa O (1990) Electrostatic manipulation of DNA in microfabricated structures. IEEE Trans Ind Appl 26:1165–1172
Tuukkanen S, Kuzyk A, Toppari JJ, Häkkinen H, Hytönen VP, Niskanen E, Rinkiö M, Törmä P (2007) Trapping of 27 bp–8 kbp DNA and immobilization of thiol-modified DNA using dielectrophoresis. Nanotechnology 18:295204. https://doi.org/10.1088/0957-4484/18/29/295204
Pohl H (1978) Dielectrophoresis. Cambridge University Press, Cambridge
Pethig R (2010) Dielectrophoresis: status of the theory, technology, and applications. Biomicrofluidics 4:022811. https://doi.org/10.1063/1.3456626
Hölzel R, Gajovic-Eichelmann N, Bier FF (2003) Oriented and vectorial immobilization of linear M13 dsDNA between interdigitated electrodes—towards single molecule DNA nanostructures. Biosens Bioelectron 18:555–564. https://doi.org/10.1016/S0956-5663(03)00024-1
Kumemura M, Collard D, Yamahata C, Sakaki N, Hashiguchi G, Fujita H (2007) Single DNA molecule isolation and trapping in a microfluidic device. ChemPhysChem 8:1875–1880. https://doi.org/10.1002/cphc.200700268
Hyun C, Kaur H, McNabb DS, Li J (2015) Dielectrophoretic stretching of DNA tethered to a fiber tip. Nanotechnology 26:125501. https://doi.org/10.1088/0957-4484/26/12/125501
Tuukkanen S, Kuzyk A, Toppari JJ, Hytönen VP, Ihalainen T, Törmä P (2005) Dielectrophoresis of nanoscale double-stranded DNA and humidity effects on its electrical conductivity. Appl Phys Lett 87:183102. https://doi.org/10.1063/1.2117626
Yamamoto T, Kurosawa O, Kabata H, Shimamoto M, Washizu M (2000) Molecular surgery of DNA based on electrostatic micromanipulation. IEEE Trans Ind Appl 36:1010–1017. https://doi.org/10.1109/IAS.1998.729860
Henning A, Henkel J, Bier FF, Hölzel R (2008) Label-free electrical quantification of the dielectrophoretic response of DNA. BMC Biophys 1:4. https://doi.org/10.1186/1757-5036-1-4
Henning A, Bier FF, Hölzel R (2010) Dielectrophoresis of DNA: quantification by impedance measurements. Biomicrofluidics 4:022803. https://doi.org/10.1063/1.3430550
Li S, Yuan Q, Morshed BI, Ke C, Wu J, Jiang H (2013) Dielectrophoretic responses of DNA and fluorophore in physiological solution by impedimetric characterization. Biosens Bioelectron 41:649–655. https://doi.org/10.1016/j.bios.2012.09.036
Stanke S, Bier FF, Hölzel R (2011) Fluid streaming above interdigitated electrodes in dielectrophoresis experiments. Electrophoresis 32:2448–2245. https://doi.org/10.1002/elps.201100096
Acknowledgments
The authors thank Katrin Nicklas for help with electrode preparations. Financial support by The Brandenburg Ministry of Sciences, Research and Cultural Affairs (MWFK) within the framework StaF and by the European Regional Development Fund (ERDF) is gratefully acknowledged. We also thank the German Research Foundation (DFG) for funding in the program Essence (SPP 1857 HO1298/4-1).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Laux, EM., Bier, F.F., Hölzel, R. (2018). Dielectrophoretic Stretching of DNA. In: Zuccheri, G. (eds) DNA Nanotechnology. Methods in Molecular Biology, vol 1811. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8582-1_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8582-1_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8581-4
Online ISBN: 978-1-4939-8582-1
eBook Packages: Springer Protocols