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Lipid-Based Passivation in Nanofluidics

[Image: see text] Stretching DNA in nanochannels is a useful tool for direct, visual studies of genomic DNA at the single molecule level. To facilitate the study of the interaction of linear DNA with proteins in nanochannels, we have implemented a highly effective passivation scheme based on lipid b...

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Autores principales: Persson, Fredrik, Fritzsche, Joachim, Mir, Kalim U., Modesti, Mauro, Westerlund, Fredrik, Tegenfeldt, Jonas O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2012
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3348678/
https://www.ncbi.nlm.nih.gov/pubmed/22432814
http://dx.doi.org/10.1021/nl204535h
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author Persson, Fredrik
Fritzsche, Joachim
Mir, Kalim U.
Modesti, Mauro
Westerlund, Fredrik
Tegenfeldt, Jonas O.
author_facet Persson, Fredrik
Fritzsche, Joachim
Mir, Kalim U.
Modesti, Mauro
Westerlund, Fredrik
Tegenfeldt, Jonas O.
author_sort Persson, Fredrik
collection PubMed
description [Image: see text] Stretching DNA in nanochannels is a useful tool for direct, visual studies of genomic DNA at the single molecule level. To facilitate the study of the interaction of linear DNA with proteins in nanochannels, we have implemented a highly effective passivation scheme based on lipid bilayers. We demonstrate virtually complete long-term passivation of nanochannel surfaces to a range of relevant reagents, including streptavidin-coated quantum dots, RecA proteins, and RecA–DNA complexes. We show that the performance of the lipid bilayer is significantly better than that of standard bovine serum albumin-based passivation. Finally, we show how the passivated devices allow us to monitor single DNA cleavage events during enzymatic degradation by DNase I. We expect that our approach will open up for detailed, systematic studies of a wide range of protein–DNA interactions with high spatial and temporal resolution.
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spelling pubmed-33486782012-05-09 Lipid-Based Passivation in Nanofluidics Persson, Fredrik Fritzsche, Joachim Mir, Kalim U. Modesti, Mauro Westerlund, Fredrik Tegenfeldt, Jonas O. Nano Lett [Image: see text] Stretching DNA in nanochannels is a useful tool for direct, visual studies of genomic DNA at the single molecule level. To facilitate the study of the interaction of linear DNA with proteins in nanochannels, we have implemented a highly effective passivation scheme based on lipid bilayers. We demonstrate virtually complete long-term passivation of nanochannel surfaces to a range of relevant reagents, including streptavidin-coated quantum dots, RecA proteins, and RecA–DNA complexes. We show that the performance of the lipid bilayer is significantly better than that of standard bovine serum albumin-based passivation. Finally, we show how the passivated devices allow us to monitor single DNA cleavage events during enzymatic degradation by DNase I. We expect that our approach will open up for detailed, systematic studies of a wide range of protein–DNA interactions with high spatial and temporal resolution. American Chemical Society 2012-03-20 2012-05-09 /pmc/articles/PMC3348678/ /pubmed/22432814 http://dx.doi.org/10.1021/nl204535h Text en Copyright © 2012 American Chemical Society http://pubs.acs.org This is an open-access article distributed under the ACS AuthorChoice Terms & Conditions. Any use of this article, must conform to the terms of that license which are available at http://pubs.acs.org.
spellingShingle Persson, Fredrik
Fritzsche, Joachim
Mir, Kalim U.
Modesti, Mauro
Westerlund, Fredrik
Tegenfeldt, Jonas O.
Lipid-Based Passivation in Nanofluidics
title Lipid-Based Passivation in Nanofluidics
title_full Lipid-Based Passivation in Nanofluidics
title_fullStr Lipid-Based Passivation in Nanofluidics
title_full_unstemmed Lipid-Based Passivation in Nanofluidics
title_short Lipid-Based Passivation in Nanofluidics
title_sort lipid-based passivation in nanofluidics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3348678/
https://www.ncbi.nlm.nih.gov/pubmed/22432814
http://dx.doi.org/10.1021/nl204535h
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AT westerlundfredrik lipidbasedpassivationinnanofluidics
AT tegenfeldtjonaso lipidbasedpassivationinnanofluidics