DNA fluorocode: A single molecule, optical map of DNA with nanometre resolution
DNA fluorocode: A single molecule, optical map of DNA with nanometre resolution
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DOI：
10.1039/c0sc00277a
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发表时间：
2010-10-01
期刊：
影响因子：
8.4
通讯作者：
Hofkens, Johan
中科院分区：
文献类型：
--
作者：
Neely, Robert K.;Dedecker, Peter;Hofkens, Johan
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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.Hhal, 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.