Use of 3'-oxymethylene alkyl disulfide protected nucleotides for enzymatic dna and rna synthesis

Abstract

This invention is about enzymatic synthesis of nucleic acids using 3′-O—(CH2SSR) protected reversible nucleotide terminators. The nucleotide base of the protected nucleotide may consist of natural or non-natural (e.g. 7-de-aza G and 7-de-aza A), or mixture thereof. The base of the nucleotides can be modified with a linker carrying carboxylic acid (—CO2H). amine (—NH2), thiol (—SH). hydroxymethyl (—CH2OH), propargy lamine. alkyne group etc. for subsequent modification and labeling. Such nucleotide substrates can be incorporated into the single strand, template free nucleic acid or into the templated DNA hybrid. And in later step the 3′-O—CH2SSR protecting group can be cleaved off by chemical treatment pre-requisites for enzymatic nucleic acids synthesis. By adding nucleotide in pre-determined fashion and cleave reaction after each step. longer DNA strands can be synthesized in solution or on solid surface starting from a short seeding DNA strands.

Dependencies are inferred from the claim text.