Enzymatic and NMR analysis of oligoribonucleotides synthesized with 2′-tert-butyldimethylsilyl protected cyanoethlphosphoramidite monomers

Yu Ying Wang, Matthew H. Lyttle, Philip N. Borer

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

The regioisomeric integrity of the internucleotide phosphate linkage in synthetic RNA using 2′-tertbutyldimethylsilyl protection was examined using enzymatic and NMR techniques. Two sets of DNA-RNA hybrid nonamers, T3XT5 and T5XT3 (where X = rA, rC, rG and U) and the tetramer AGCU were analyzed. Enzyme catalyzed hydrolysis of the nonamers with ribonuclease T2 showed that the linkage at the ribonucleotide was the desired 3′-5′. A control nonamer with a 2′-5′ linkage was subjected to the enzyme, and showed no cleavage. High-resolution proton NMR of the tetramer also gave a favorable comparison with the same molecule obtained by non-chemical means.

Original languageEnglish (US)
Pages (from-to)3347-3352
Number of pages6
JournalNucleic acids research
Volume18
Issue number11
DOIs
StatePublished - Jun 11 1990

ASJC Scopus subject areas

  • Genetics

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