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Updated: Aug 23, 2025

Robust 3D DNA FISH Using Directly Labeled Probes
Published on: August 15, 2013
A new phosphoramidite enables orthogonal double labelling to form combination oligonucleotide probes
Chunsen Bai1, Piotr Klimkowski1, Cheng Jin1
1Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, UK. tom.brown@chem.ox.ac.uk.
Abstract:
Oligonucleotides labelled with thiazole orange intercalator and a reporter dye on the same thymine base have been synthesized. The key phosphoramidite (AP-C3 dT) contains an alkyne and amine, enabling dual orthogonal labelling of the nucleobase. Multiple monomers can be added to produce heavily functionalised oligonucleotides. In their DNA and 2'-OMe RNA formats these combination probes display high duplex stability and fluorescence when bound to complementary DNA and RNA.
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