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Microwave-Mediated Synthesis of Labeled Nucleotides with Utility in the Synthesis of DNA Probes
Journal article   Peer reviewed

Microwave-Mediated Synthesis of Labeled Nucleotides with Utility in the Synthesis of DNA Probes

Mark Lefever, Jerome W. Kosmeder, Michael Farrell, Christopher Bieniarz and Christopher B Rodell
Bioconjugate chemistry, v 21(10), pp 1773-1778
01 Oct 2010
PMID: 20812713

Abstract

Biochemical Research Methods Biochemistry & Molecular Biology Chemistry Chemistry, Multidisciplinary Chemistry, Organic Life Sciences & Biomedicine Physical Sciences Science & Technology
A novel method of linking haptens to deoxycytidine 5'-triphosphate via microwave-mediated bisulfate-catalyzed transamination with hydrazine has been developed. This method enables the tethering of small molecule haptens to dCTP via a discrete polyethylene glycol (PEG) spacer, yielding N-4-aminodeoxycytidine 5'-triphosphate-dPEG-haptens. This synthetic approach employs microwave-catalyzed hydrazinolysis that enables the attachment of spacers via hydrazine linkages. The microwave-mediated introduction of this hydrazine handle provides a significant improvement in yield over those of published thermal methods. The microwave reaction was shown to be scalable, and the final product was amenable to labeling with a wide variety of haptens. The resulting nucleotide triphosphates, N-4-aminodeoxycytidine 5'-triphosphate-dPEG-haptens, can serve as unique substrates for the enzyme-mediated labeling of DNA probes. The efficacy of incorporation of one such novel nucleotide, N-4-aminodeoxycyticline 5'-triphosphate-dPEG(4)-DNP, has been demonstrated in nick translation labeling of HER2 and HPV probes. The labeled probes have been shown to be effective in visualizing their target genes in tissue.

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Web of Science research areas
Biochemical Research Methods
Biochemistry & Molecular Biology
Chemistry, Multidisciplinary
Chemistry, Organic
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