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Nucleic Acids Research, 1990, Vol. 18, No. 11 3347-3352
© 1990

CHEMISTRY

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

Yu Ying Wang, Matthew H. Lyttle1 and Philip N. Borer*

Department of Chemistry, Syracuse University Syracuse, Ny 13244 1MilliGen/Biosearch, Division of Millipore 81 Digital Drive, Novato, CA 94949, USA

*To whom correspondence should be addressed

Received December 29, 1989. Accepted April 24, 1990.

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.


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