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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Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52
Geninatti Crich S.
Alberti D.
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Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52
Alberti D.
Orio L.
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Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52
Orio L.
Stefania R.
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Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52
Stefania R.
Longo D.
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Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52
Longo D.
Aime S.
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Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52Department of Molecular Biotechnology and Health Sciences, University of Torino, via Nizza 52
机构:
Univ Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
Univ Massachusetts, Dept Biomed Engn, Amherst, MA 01003 USAUniv Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
Ramesh, Anujan
Kumar, Sahana
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Univ Massachusetts, Dept Chem Engn, Amherst, MA 01003 USAUniv Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
Kumar, Sahana
Nguyen, Anh
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Univ Massachusetts, Dept Chem Engn, Amherst, MA 01003 USAUniv Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
Nguyen, Anh
Brouillard, Anthony
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Univ Massachusetts, Dept Chem Engn, Amherst, MA 01003 USAUniv Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
Brouillard, Anthony
Kulkarni, Ashish
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Univ Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
Univ Massachusetts, Dept Biomed Engn, Amherst, MA 01003 USA
Univ Massachusetts, Ctr Bioact Delivery, Inst Appl Life Sci, Amherst, MA 01003 USAUniv Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
机构:
Univ Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, Italy
Univ Roma La Sapienza, Dept Expt Med, Sect Med Physiopathol Endocrinol & Nutr, Li Sa Lab, Rome, ItalyUniv Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, Italy
Carbone, Claudia
Leonardi, Antonio
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Univ Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, ItalyUniv Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, Italy
Leonardi, Antonio
Cupri, Sarha
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Univ Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, ItalyUniv Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, Italy
Cupri, Sarha
Puglisi, Giovanni
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Univ Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, ItalyUniv Catania, Dept Drug Sci, Sect Pharmaceut Technol, Catania, Italy
机构:
Univ N Carolina, Div Mol Pharmaceut, Eshelman Sch Pharm, Chapel Hill, NC 27599 USAUniv N Carolina, Div Mol Pharmaceut, Eshelman Sch Pharm, Chapel Hill, NC 27599 USA
Huang, Leaf
Liu, Yang
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Univ N Carolina, Div Mol Pharmaceut, Eshelman Sch Pharm, Chapel Hill, NC 27599 USAUniv N Carolina, Div Mol Pharmaceut, Eshelman Sch Pharm, Chapel Hill, NC 27599 USA
机构:Univ Calif San Francisco, Sch Pharm, Dept Biopharmaceut Sci, San Francisco, CA 94143 USA
Li, Weijun
Szoka, Francis C., Jr.
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Univ Calif San Francisco, Sch Pharm, Dept Biopharmaceut Sci, San Francisco, CA 94143 USAUniv Calif San Francisco, Sch Pharm, Dept Biopharmaceut Sci, San Francisco, CA 94143 USA