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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2012
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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. |
format | Online Article Text |
id | pubmed-3348678 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT perssonfredrik lipidbasedpassivationinnanofluidics AT fritzschejoachim lipidbasedpassivationinnanofluidics AT mirkalimu lipidbasedpassivationinnanofluidics AT modestimauro lipidbasedpassivationinnanofluidics AT westerlundfredrik lipidbasedpassivationinnanofluidics AT tegenfeldtjonaso lipidbasedpassivationinnanofluidics |