title: Simple, High-Yield Syntheses of DNA Duplexes Containing Interstrand DNA-DNA Cross-Links Between an N(4) -Aminocytidine Residue and an Abasic Site.
abstract: The protocol describes the preparation and purification of interstrand DNA-DNA cross-links derived from the reaction of an N(4) -aminocytidine residue with an abasic site in duplex DNA. The procedures employ inexpensive, commercially available chemicals and enzymes to carry out post-synthetic modification of commercially available oligodeoxynucleotides. The yield of cross-linked duplex is typically better than 90%. If purification is required, the cross-linked duplex can be readily separated from single-stranded DNA starting materials by denaturing gel electrophoresis. The resulting covalent hydrazone-based cross-links are stable under physiologically relevant conditions and may be useful for biophysical studies, structural analyses, DNA repair studies, and materials science applications. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Department of Chemistry, University of Missouri, Columbia, Missouri.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Gamboa Varela
authors.affiliation: Department of Chemistry, University of Missouri, Columbia, Missouri.|Department of Biochemistry, University of Missouri, Columbia, Missouri.
authors.forename: Kent S
authors.initials: KS
authors.lastname: Gates
mesh_terms: D060326:Chemistry Techniques, Synthetic; D003432:Cross-Linking Reagents / Q000737:chemistry; D003562:Cytidine / Q000737:chemistry*; D004247:DNA / Q000737:chemistry; D063347:Native Polyacrylamide Gel Electrophoresis; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D010761:Phosphorus Radioisotopes / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides; D010761:Phosphorus Radioisotopes; D003562:Cytidine; D004247:DNA
keywords: DNA cross-link; DNA repair; abasic; click chemistry; hydrazone; nanomaterials; oligonucleotide