A micropatterned substrate for on-surface enzymatic labelling of linearized long DNA molecules View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2019-10-21

AUTHORS

Dharma Varapula, Eric LaBouff, Kaitlin Raseley, Lahari Uppuluri, Garth D. Ehrlich, Moses Noh, Ming Xiao

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. More... »

PAGES

15059

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41598-019-51507-z

DOI

http://dx.doi.org/10.1038/s41598-019-51507-z

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1121973438

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/31636335


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