Oligonucleotide dendrimers: Synthesis and use as polylabelled DNA probes

• Published in: Nucleic acids research Vol. 25; no. 22; pp. 4447 - 4454
• Main Authors: Shchepinov, M. S., Udalova, I. A., Bridgman, A. J., Southern, E. M.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 15.11.1997
• Subjects: Adenosine Triphosphate; Biopolymers; DNA Primers - chemical synthesis; DNA Probes - chemical synthesis; Isotope Labeling; Oligonucleotides - chemical synthesis; Phosphorus Radioisotopes; Polymerase Chain Reaction; Polynucleotide 5'-Hydroxyl-Kinase
• ISSN: 0305-1048; 1362-4962; 1362-4962
• DOI: 10.1093/nar/25.22.4447

Oligonucleotide dendrimers were synthesized using a novel phosphoramidite synthon, tris-2,2,2-[3-(4,4′-dimethoxytrityloxy) propyloxymethyl]ethyl-N,N-diisopropylaminocyanethoxy phosphoramidite. Label, incorporated using [γ-32P]ATP and polynucleotide kinase, was increased in proportion to the number of 5′-ends. There was a similar increase in signal when these multiply labelled oligonucleotides were used as probes to oligonucleotide arrays. A dendrimeric oligonucleotide was used successfully as a primer in the PCR. The strand bearing the dendrimer was resistant to degradation by T7 Gene 6 exonuclease making it easy to convert the double-stranded product of the PCR to a multiply-labelled, single-stranded probe.