1] Chemical synthesis of deoxyoligonucleotides and deoxyoligonucleotide analogs

Abstract

Publisher Summary This chapter discusses advances in chemical methodologies, which include the development of new protecting group strategies and procedures for the attachment of reporter groups to DNA. Because of these achievements, the time required to synthesize DNA has been reduced, and an even larger repertoire of applications for synthetic deoxyoligonucleotides has been achieved. An attempt is made to review some of these many excellent contributions, but only one detailed protocol for labeling DNA is included. This is a relatively recent, unpublished procedure for attaching reporter groups to a new DNA analog called “dithioate DNA.” Deoxyoligonucleotides having a deoxynucleoside–OPS 2 O–deoxynucleoside linkage, dithioate DNA, are excellent mimics of natural DNA. At least partially, this is because they are isosteric and isoelectronic with normal DNA. They are, however, completely resistant to snake venom, calf spleen, P 1 exonucleases, all the nucleases found in HeLa nuclear and cytoplasmic extracts, and even the very potent exonucleolytic activity of T 4 DNA polymerase. These attractive features combined with the ability of dithioate DNA to form stable DNA duplexes have led to several potential applications.