Single-Stranded DNA Curtains for Real-Time Single-Molecule Visualization of Protein–Nucleic Acid Interactions
Year: 2012
Type: article
Abstract: Single-molecule imaging of biological macromolecules has dramatically impacted our understanding of many types of biochemical reactions. To facilitate these studies, we have established new strategies for anchoring and organizing DNA molecules on the surfaces of microfluidic sample chambers that are otherwise coated with fluid lipid bilayers. This previous work was reliant upon the use of double-stranded DNA, precluding access to information on biological processes involving single-stranded nucleic acid substrates. Here, we present procedures for aligning and visualizing single-stranded DNA molecules along the leading edges of nanofabricated barriers to lipid diffusion, in both "single-tethered" and "double-tethered" experimental formats. This new single-molecule approach provides long-awaited access to critical biological reactions involving single-stranded DNA binding proteins.
Source: Analytical Chemistry
DOI: 10.1021/ac302117z
Institutions: Howard Hughes Medical Institute, Columbia University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: bronze
Awards: G2505422825, Elucidating the Mechanisms of DNA Recombination, Visualizing the Dynamics of Chromatin and Chromatin Remodeling Proteins, Nanoscience and Engineering Center: Columbia Center for Electronic Transport in Molecular Nanostructures, Using nanofabricated DNA curtains to reveal protein during replication, G8567027302, G8875402815
OpenAlex ID: W2016225119