Site-specific DNA labelling by Staudinger ligation

Weisbrod, Samuel H.; Baccaro, Anna; Marx, Andreas

Site-specific DNA labelling by Staudinger ligation

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Date

2011

Authors

Weisbrod, Samuel H.

Baccaro, Anna

Marx, Andreas

Editors

Walker, John

Publication type

Contribution to a collection

Published in

Bioconjugation Protocols / Mark, Sonny S. (ed.). - Totowa, NJ : Humana Press, 2011. - (Methods in Molecular Biology ; 751). - pp. 195-207. - ISBN 978-1-61779-150-5

Abstract

Site-specific and chemoselective labeling of DNA is still a difficult task. The Staudinger ligation is a bioorthogonal reaction between azides and phosphines that requires no catalyst to proceed, allowing for mild reaction conditions. The reaction may be extended for site-specific labeling of DNA using azidomodified triphosphates, which can be incorporated site-specifically into DNA strands by DNA polymerases in a template-dependent manner. The azido-modified DNA, in turn, can be labeled by suitable phosphines. This protocol describes (1) the synthesis of an azido-TTP analogue; (2) the enzymatic synthesis of azido-modified DNA; (3) the synthesis of suitable phosphine labels; and (4) the labeling of azido-DNA with biotin–phosphine by Staudinger ligation with approximately 70% conversion.

Subject (DDC)

540 Chemistry

Keywords

Site-specific DNA labeling,DNA modification,Staudinger ligation,Phosphine,Modified triphosphate,Azido thymidine,DNA polymerase,Azido-DNA,Biotin

Cite This

ISO 690

WEISBROD, Samuel H., Anna BACCARO, Andreas MARX, WALKER, John, ed., 2011. Site-specific DNA labelling by Staudinger ligation. In: MARK, Sonny S., ed.. Bioconjugation Protocols. Totowa, NJ:Humana Press, pp. 195-207. ISBN 978-1-61779-150-5. Available under: doi: 10.1007/978-1-61779-151-2_12

BibTex

@incollection{Weisbrod2011Sites-14013,
  year={2011},
  doi={10.1007/978-1-61779-151-2_12},
  title={Site-specific DNA labelling by Staudinger ligation},
  number={751},
  isbn={978-1-61779-150-5},
  publisher={Humana Press},
  address={Totowa, NJ},
  series={Methods in Molecular Biology},
  booktitle={Bioconjugation Protocols},
  pages={195--207},
  editor={Mark, Sonny S.},
  author={Weisbrod, Samuel H. and Baccaro, Anna and Marx, Andreas}
}

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