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Synthesis and oligonucleotide incorporation of fluorescent cytosine analogue tC: a promising nucleic acid probe

Nature Protocols volume 2, pages 615–623 (2007)Cite this article

Abstract

The tricyclic cytosine, tC, is a fluorescent base analogue with excellent properties for investigating intrinsic characteristics of nucleic acid as well as interactions between nucleic acids and other molecules. Its unique fluorescence properties and insignificant influence on overall structure and dynamics of nucleic acid after incorporation makes tC particularly interesting in fluorescence resonance energy transfer and anisotropy measurements. We here describe a straightforward synthesis of the standard monomer form of tC for DNA solid-phase synthesis, the tC phosphoramidite, and its subsequent incorporation into oligonucleotides. The total synthesis of the tC phosphoramidite takes approximately 8 days and its incorporation and the subsequent oligonucleotide purification an additional day.

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Authors and Affiliations

  1. Department of Chemical and Biological Engineering/Physical Chemistry, Chalmers University of Technology, Gothenburg, SE-41296, Sweden

    Peter Sandin, Per Lincoln & L Marcus Wilhelmsson

  2. School of Chemistry, University of Southampton, Highfield, SO17 1BJ, Southampton, UK

    Tom Brown

Authors
  1. Peter Sandin
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  2. Per Lincoln
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  3. Tom Brown
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  4. L Marcus Wilhelmsson
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Correspondence to L Marcus Wilhelmsson.

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Sandin, P., Lincoln, P., Brown, T. et al. Synthesis and oligonucleotide incorporation of fluorescent cytosine analogue tC: a promising nucleic acid probe. Nat Protoc 2, 615–623 (2007). https://doi.org/10.1038/nprot.2007.80

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