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Issue 4, 2010
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DNA fluorocode: A single molecule, optical map of DNA with nanometre resolution

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Abstract

We present a new method for single-molecule optical DNA mapping using an exceptionally dense, yet sequence-specific coverage of DNA with a fluorescent probe. The method employs a DNA methyltransferase enzyme to direct the DNA labelling, followed by molecular combing of the DNA onto a polymer-coated surface and subsequent sub-diffraction limit localization of the fluorophores. The result is a ‘DNA fluorocode’; a simple description of the DNA sequence, with a maximum achievable resolution of less than 20 bases, which can be read and analyzed like a barcode. We demonstrate the generation of a fluorocode for genomic DNA from the lambda bacteriophage using a DNA methyltransferase, M.HhaI, to direct fluorescent labels to four-base sequences reading 5′-GCGC-3′. A consensus fluorocode that allows the study of the DNA sequence at the level of an individual labelling site can be generated from a handful of molecules.

Graphical abstract: DNA fluorocode: A single molecule, optical map of DNA with nanometre resolution

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Supplementary files

Article information


Submitted
22 Apr 2010
Accepted
05 Jul 2010
First published
11 Aug 2010

Chem. Sci., 2010,1, 453-460
Article type
Edge Article

DNA fluorocode: A single molecule, optical map of DNA with nanometre resolution

R. K. Neely, P. Dedecker, J. Hotta, G. Urbanavičiūtė, S. Klimašauskas and J. Hofkens, Chem. Sci., 2010, 1, 453
DOI: 10.1039/C0SC00277A

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