Open AccessSimple, High‐Yield Syntheses of DNA Duplexes Containing Interstrand DNA‐DNA Cross‐Links Between an N 4 ‐Aminocytidine Residue and an Abasic Site
Author(s) -
Gamboa Varela Jacqueline,
Gates Kent S.
Publication year - 2016
Publication title -
current protocols in nucleic acid chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.306
H-Index - 17
eISSN - 1934-9289
pISSN - 1934-9270
DOI - 10.1002/cpnc.3
Subject(s) - dna , duplex (building) , chemistry , residue (chemistry) , dna ligase , combinatorial chemistry , covalent bond , ap site , yield (engineering) , biochemistry , dna repair , organic chemistry , materials science , metallurgy
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.