A Large-Scale Synthesis of a Nonadecadeoxyribonucleotide Duplex Having a Sequence Identical to That of Phage φ80 O_R2 by the Phosphoro-p-anisidate Method

Abstract

A nonadecanucleotide duplex, dCATCACCATAATGTTCATT·dAATGAACATTATGGTGATG, having a sequence identical to that of phage φ80 O_R2 has been synthesized by the phosphoro-p-anisidate method. 5'-O-Dimethoxytrityl-N-protected deoxynucleotide 3'-(o-chlorophenyl)phosphates and N-protected deoxynucleoside 3'-(o-chlorophenyl)phosphoro-p-anisidates were used to prepare di- and trinucleotide blocks. Oligomers were elongated either in the 5'-direction by dedimethoxytritylation or in the 3'-direction by treatment with isoamyl nitrite, and isolated by chromatography on silica gel or alkylated silica gel. The deblocked nonadecamers were purified by reversed phase and ion-exchange chromatography and the duplex was isolated by gel filtration.