Highly efficient chemical synthesis of 2'-O-methyl-oligoribonucleotides and tetrabiotinylated derivatives; novel probes that are resistant to degradation by RNA or DNA specific nucleases

2'-O-methyl-oligoribonucleotides have been synthesised on solid phase from base protected 5'-O-dimethoxytrityl-2'-O-methylribonucleoside-3'-O-(2-cyanoethyl N,N-diisopropyl-phosphoramidites) using 5-(4-nitrophenyl)-1H-tetrazole as activator. Coupling yields greater than 99% were a...

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Bibliographic Details
Published in:Nucleic acids research 1989-01, Vol.17 (9), p.3373-3386
Main Authors: Sproat, B S, Lamond, AI, Beijer, B, Neuner, P, Ryder, U
Format: Article
Language:English
Online Access:Get full text
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Summary:2'-O-methyl-oligoribonucleotides have been synthesised on solid phase from base protected 5'-O-dimethoxytrityl-2'-O-methylribonucleoside-3'-O-(2-cyanoethyl N,N-diisopropyl-phosphoramidites) using 5-(4-nitrophenyl)-1H-tetrazole as activator. Coupling yields greater than 99% were achieved, as judged by trityl cation release. The preparation of a modified 2'-deoxycytidine building block bearing and N super(4)-(5-trifluoroacetyl-aminopentyl) spacer is also described. The latter compound enabled the chemical synthesis of 2'-O-methyl-oligoribonucleotide probes carrying several 5'-terminal biotinylation sites (in general four modified residues were used), which can be conveniently super(32)P end-labelled enzymatically using polynucleotide kinase.
ISSN:0305-1048