Title: Simple, High-Yield Syntheses of DNA Duplexes Containing Interstrand DNA-DNA Cross-Links Between an N(4) -Aminocytidine Residue and an Abasic Site.
Authors: Gamboa Varela, Jacqueline; Gates, Kent S
Published In Curr Protoc Nucleic Acid Chem, (2016 06 01)
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.
PubMed ID: 27248783
MeSH Terms: Chemistry Techniques, Synthetic; Cross-Linking Reagents/chemistry; Cytidine/chemistry*; DNA/chemistry; Native Polyacrylamide Gel Electrophoresis; Nucleic Acid Heteroduplexes/chemistry*; Oligodeoxyribonucleotides/chemical synthesis; Oligodeoxyribonucleotides/chemistry*; Phosphorus Radioisotopes/chemistry