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Issue 21, 2013
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Protein–DNA force assay in a microfluidic format

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Abstract

The detailed study of protein–DNA interactions is a core effort to elucidate physiological processes, including gene regulation, DNA repair and the immune response. The molecular force assay (MFA) is an established method to study DNA-binding proteins. In particular, high-affinity binder dissociation is made possible by the application of force. Microfluidic lab-on-a-chip approaches have proven helpful for parallelization, small sample volumes, reproducibility, and low cost. We report the successful combination of these two principles, forming a microfluidic molecular force assay and representing a novel use for the established MITOMI chip design. We present, characterize, validate and apply this integrated method. An alternative confocal fluorescence microscopy readout and analysis method is introduced and validated. In a multiplexing application, EcoRI binding is detected and characterized. This method paves the way for quantitative on-chip force measurements. It is suited for integration with DNA micro-spotting and in vitro expression of transcription factors to form a high-throughput chip for detailed DNA–protein interaction studies.

Graphical abstract: Protein–DNA force assay in a microfluidic format

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Supplementary files

Article information


Submitted
12 Jul 2013
Accepted
02 Aug 2013
First published
29 Aug 2013

Lab Chip, 2013,13, 4198-4204
Article type
Paper

Protein–DNA force assay in a microfluidic format

M. Otten, P. Wolf and H. E. Gaub, Lab Chip, 2013, 13, 4198
DOI: 10.1039/C3LC50830G

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