In order to synthesize oligonucleotide, bifunctional agent, bis-(N, N-dialkylamino) methoxyphosphine and stable monofunctional agent, deoxynucleoside-morpholino-methoxy-phosphine have been synthesized through modified one-bottle method and utilized in manual synthesis and automatic synthesis with Beckman System one plus DNA synthesizer. In the case of bis-(N,N-dialkylamino) methoxyphosphine, the density of functional group on solid-surface and the purity of solvent and reagent appear to be an important factor to avoid of interchain cross-linking and impurity amplication.
Ultrasonic effect for deprotection of synthetic oligonucleotide from solid support was investigated with comparision of standard condition. The rate of deprotection was remarkably increased. Therefore the effect of ultrasound was proved to be a good accelator of releasing reaction from solid support.
Single strand circular DNA encoding sequence of lac promotor has been synthesized first with 41 base long single strand linear DNA synthesized by automatic synthesizer. The product was characterized on polyacrylamide gel in terms of gel mobilities of linear and circularized DNA fragment obtained from EcoRI digestion.
