title: Use of a novel 5'-regioselective phosphitylating reagent for one-pot synthesis of nucleoside 5'-triphosphates from unprotected nucleosides.
abstract: 5'-Triphosphates are building blocks for enzymatic synthesis of DNA and RNA. This unit presents a protocol for convenient synthesis of 2'-deoxyribo- and ribonucleoside 5'-triphosphates (dNTPs and NTPs) from any natural or modified base. This one-pot synthesis can also be employed to prepare triphosphate analogs with a sulfur or selenium atom in place of a non-bridging oxygen atom of the α-phosphate. These S- or Se-modified dNTPs and NTPs can be used to prepare diastereomerically pure phosphorothioate or phosphoroselenoate nucleic acids. Even without extensive purification, the dNTPs or NTPs synthesized by this method are of high quality and can be used directly in DNA polymerization or RNA transcription. Synthesis and purification of the 5'-triphosphates, as well as analysis and confirmation of natural and sulfur- or selenium-modified nucleic acids, are described in this protocol unit.
authors.affiliation: Department of Chemistry and Department of Biology, Georgia State University, Atlanta, Georgia.
authors.forename: Julianne
authors.initials: J
authors.lastname: Caton-Williams
authors.affiliation: Department of Chemistry and Department of Biology, Georgia State University, Atlanta, Georgia.
authors.forename: Rudiona
authors.initials: R
authors.lastname: Hoxhaj
authors.affiliation: Department of Chemistry and Department of Biology, Georgia State University, Atlanta, Georgia.
authors.forename: Bilal
authors.initials: B
authors.lastname: Fiaz
authors.affiliation: Department of Chemistry and Department of Biology, Georgia State University, Atlanta, Georgia.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Huang
mesh_terms: D002625:Chemistry, Organic / Q000379:methods*; D009705:Nucleosides / Q000138:chemical synthesis / Q000737:chemistry*; D055601:Organic Chemistry Phenomena; D010084:Oxidation-Reduction; D010710:Phosphates / Q000737:chemistry; D011122:Polyphosphates / Q000737:chemistry; D018036:Selenium Compounds / Q000737:chemistry
chemical_list: D009705:Nucleosides; D010710:Phosphates; D011122:Polyphosphates; D018036:Selenium Compounds; C005692:triphosphoric acid; C035638:thiophosphoric acid