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• W2027287466 abstract "Abstract We demonstrate an approach to optical DNA mapping, which enables near single-molecule characterization of whole bacteriophage genomes. Our approach uses a DNA methyltransferase enzyme to target labelling to specific sites and copper-catalysed azide-alkyne cycloaddition to couple a fluorophore to the DNA. We achieve a labelling efficiency of ∼70% with an average labelling density approaching one site every 500 bp. Such labelling density bridges the gap between the output of a typical DNA sequencing experiment and the long-range information derived from traditional optical DNA mapping. We lay the foundations for a wider-scale adoption of DNA mapping by screening 11 methyltransferases for their ability to direct sequence-specific DNA transalkylation; the first step of the DNA labelling process and by optimizing reaction conditions for fluorophore coupling via a click reaction. Three of 11 enzymes transalkylate DNA with the cofactor we tested (a readily prepared s-adenosyl-l-methionine analogue)." @default.
• W2027287466 created "2016-06-24" @default.
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• W2027287466 date "2014-01-21" @default.
• W2027287466 modified "2023-10-16" @default.
• W2027287466 title "Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry" @default.
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• W2027287466 doi "https://doi.org/10.1093/nar/gkt1406" @default.
• W2027287466 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3985630" @default.
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