Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry. / Vranken, Charlotte; Deen, Jochem; Dirix, Lieve; Stakenborg, Tim; Dehaen, Wim; Leen, Volker; Hofkens, Johan; Neely, Robert K.

In: Nucleic Acids Research, Vol. 42, No. 7, e50, 01.04.2014.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Vranken, C, Deen, J, Dirix, L, Stakenborg, T, Dehaen, W, Leen, V, Hofkens, J & Neely, RK 2014, 'Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry', Nucleic Acids Research, vol. 42, no. 7, e50. https://doi.org/10.1093/nar/gkt1406

APA

Vranken, C., Deen, J., Dirix, L., Stakenborg, T., Dehaen, W., Leen, V., Hofkens, J., & Neely, R. K. (2014). Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry. Nucleic Acids Research, 42(7), [e50]. https://doi.org/10.1093/nar/gkt1406

Vancouver

Vranken C, Deen J, Dirix L, Stakenborg T, Dehaen W, Leen V et al. Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry. Nucleic Acids Research. 2014 Apr 1;42(7). e50. https://doi.org/10.1093/nar/gkt1406

Author

Vranken, Charlotte ; Deen, Jochem ; Dirix, Lieve ; Stakenborg, Tim ; Dehaen, Wim ; Leen, Volker ; Hofkens, Johan ; Neely, Robert K. / Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry. In: Nucleic Acids Research. 2014 ; Vol. 42, No. 7.

Bibtex

@article{f2c5b7177c4b49529457f8d8b976e597,
title = "Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry",
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). ",
author = "Charlotte Vranken and Jochem Deen and Lieve Dirix and Tim Stakenborg and Wim Dehaen and Volker Leen and Johan Hofkens and Neely, {Robert K.}",
year = "2014",
month = apr,
day = "1",
doi = "10.1093/nar/gkt1406",
language = "English",
volume = "42",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "7",

}

RIS

TY - JOUR

T1 - Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry

AU - Vranken, Charlotte

AU - Deen, Jochem

AU - Dirix, Lieve

AU - Stakenborg, Tim

AU - Dehaen, Wim

AU - Leen, Volker

AU - Hofkens, Johan

AU - Neely, Robert K.

PY - 2014/4/1

Y1 - 2014/4/1

N2 - 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).

AB - 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).

U2 - 10.1093/nar/gkt1406

DO - 10.1093/nar/gkt1406

M3 - Journal article

C2 - 24452797

VL - 42

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 7

M1 - e50

ER -

ID: 140026157