Lipid-based passivation in nanofluidics.

Nano letters (2012-03-22)
Fredrik Persson, Joachim Fritzsche, Kalim U Mir, Mauro Modesti, Fredrik Westerlund, Jonas O Tegenfeldt
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Atto 488 NHS ester, BioReagent, suitable for fluorescence, ≥80% (coupling to amines)
Sigma-Aldrich
Atto 488, suitable for fluorescence, ≥90% (HPCE)
Sigma-Aldrich
Atto 488-Biotin, BioReagent, suitable for fluorescence, ~75% (HPCE)
Sigma-Aldrich
Atto 488 maleimide, BioReagent, suitable for fluorescence, ≥90% (coupling to thiols)
Sigma-Aldrich
Atto 488 amine
Sigma-Aldrich
Atto 488 azide
Sigma-Aldrich
Atto 488 iodoacetamide

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