Polymerase Synthesis of Base-Modified DNA

Abstract

Enzymic synthesis of base-modified DNA by polymerase incorporation of modified nucleotides is discussed. Modified 2'-deoxyribonucleoside triphosphates (dNTPs) are key substrates for polymerases and can be prepd. either by triphosphorylation of modified nucleosides or by direct aq. cross-coupling reactions of halogenated dNTPs with alkynes, arylboronic acids, or alkenes. The methods of polymerase synthesis include primer extension, PCR, nicking enzyme amplifications, and other methods which enable the synthesis of diverse types of long or short and double-stranded DNA or single-stranded oligonucleotides. The applications include labeling in diagnostics (labeling or coding of DNA bases) and chem. biol. (bioconjugations, modulation of protein binding, etc.). [on SciFinder(R)]