Template and primer based synthesis of enzymatically cleavab le oligonucleotides

摘要

The present invention relates to methods of enzymatically synthesizing chirally pure oligonucleotides. In particular, it relates to methods of enzymatically synthesizing oligonucleotides using optionally digestible templates which are capable of directing the synthesis of an oligonudleotide product which can optionally be cleaved into two or more individual oligonucleotides, optionally using 3' -ribonucleotide primers. In this manner, more than one oligonucleotide having the same or different length and/or sequence can be simultaneously synthesized The oligonucleotides optionally have at least one phosphorothioate linkage, and contain certain restriction endonuclease restriction sites herein identified as cleavable within phosphorothioate oligonucleotides, thus facilitating the separation and purification of the oligonucleotides after synthesis.