High-Throughput Universal DNA Curtain Arrays for Single-Molecule Fluorescence Imaging

Langmuir : the ACS Journal of Surfaces and Colloids
Ignacio F GallardoIlya J Finkelstein

Abstract

Single-molecule studies of protein-DNA interactions have shed critical insights into the molecular mechanisms of nearly every aspect of DNA metabolism. The development of DNA curtains-a method for organizing arrays of DNA molecules on a fluid lipid bilayer-has greatly facilitated these studies by increasing the number of reactions that can be observed in a single experiment. However, the utility of DNA curtains is limited by the challenges associated with depositing nanometer-scale lipid diffusion barriers onto quartz microscope slides. Here, we describe a UV lithography-based method for large-scale fabrication of chromium (Cr) features and organization of DNA molecules at these features for high-throughput single-molecule studies. We demonstrate this approach by assembling 792 independent DNA arrays (containing >900,000 DNA molecules) within a single microfluidic flowcell. As a first proof of principle, we track the diffusion of Mlh1-Mlh3-a heterodimeric complex that participates in DNA mismatch repair and meiotic recombination. To further highlight the utility of this approach, we demonstrate a two-lane flowcell that facilitates concurrent experiments on different DNA substrates. Our technique greatly reduces the challenges a...Continue Reading

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Citations

Feb 4, 2016·Nature Communications·Maxwell W BrownIlya J Finkelstein
Aug 9, 2016·Progress in Biophysics and Molecular Biology·Logan R Myler, Ilya J Finkelstein
Jun 18, 2017·Biochemical Society Transactions·Flynn R HillAntoine M van Oijen
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May 20, 2017·Scientific Reports·Yoori KimIlya J Finkelstein
Jul 6, 2019·Nature Communications·Marie-Christine CaronJean-Yves Masson
Nov 30, 2019·Science·Yoori KimHongtao Yu
Jul 6, 2020·Nucleic Acids Research·Hongshan ZhangIlya J Finkelstein
Nov 30, 2018·Proceedings of the National Academy of Sciences of the United States of America·Sudipta BhattacharyyaRasika M Harshey
Mar 7, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Shunsuke TakahashiShinji Katsura
Mar 11, 2021·Langmuir : the ACS Journal of Surfaces and Colloids·Aurimas Kopu StasMarijonas Tutkus
Mar 18, 2021·Nano Letters·Carlos Calcines-CruzArmando Hernandez-Garcia
Jun 6, 2021·Proceedings of the National Academy of Sciences of the United States of America·Jingqi DaiJean-Baptiste Charbonnier
Jul 26, 2018·Langmuir : the ACS Journal of Surfaces and Colloids·Jeffrey M SchaubIlya J Finkelstein
Jun 4, 2019·Molecular Cell·Michael M SoniatIlya J Finkelstein

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