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Journal article
A micropatterned substrate for on-surface enzymatic labelling of linearized long DNA molecules
Scientific reports, v 9(1), pp 15059-11
21 Oct 2019
PMID: 31636335
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Optical mapping of linearized DNA molecules is a promising new technology for sequence assembly and scaffolding, large structural variant detection, and diagnostics. This is currently achieved either using nanochannel confinement or by stretching single DNA molecules on a solid surface. While the first method necessitates DNA labelling before linearization, the latter allows for modification post-linearization, thereby affording increased process flexibility. Each method is constrained by various physical and chemical limitations. One of the most common techniques for linearization of DNA uses a hydrophobic surface and a receding meniscus, termed molecular combing. Here, we report the development of a microfabricated surface that can not only comb the DNA molecules efficiently but also provides for sequence-specific enzymatic fluorescent DNA labelling. By modifying a glass surface with two contrasting functionalities, such that DNA binds selectively to one of the two regions, we can control DNA extension, which is known to be critical for sequence-recognition by an enzyme. Moreover, the surface modification provides enzymatic access to the DNA backbone, as well as minimizing non-specific fluorescent dye adsorption. These enhancements make the designed surface suitable for large-scale and high-resolution single DNA molecule studies.
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Details
- Title
- A micropatterned substrate for on-surface enzymatic labelling of linearized long DNA molecules
- Creators
- Dharma Varapula - Drexel UniversityEric LaBouff - Drexel UniversityKaitlin Raseley - Drexel UniversityLahari Uppuluri - Drexel UniversityGarth D Ehrlich - Drexel UniversityMoses Noh - Drexel UniversityMing Xiao - Drexel University
- Publication Details
- Scientific reports, v 9(1), pp 15059-11
- Publisher
- Springer Nature
- Grant note
- R01 HG005946 / NHGRI NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology; School of Biomedical Engineering, Science, and Health Systems; Mechanical Engineering and Mechanics
- Web of Science ID
- WOS:000491226200042
- Scopus ID
- 2-s2.0-85073656904
- Other Identifier
- 991019169691204721
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- Web of Science research areas
- Biochemistry & Molecular Biology