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Nucleic Acids Research, 1993, Vol. 21, No. 7 1563-1568
© 1993

METHODS

A novel method for site specific introduction of single model oxidative DNA lesions into oligodeoxyribonucleotides

Zafer Hatahet, Andrei A. Purmal and Susan S. Wallace*

Department of Microbiology and Molecular Genetics and The Markey Center for Molecular Genetics, University of Vermont Burlington, VT 05405, USA

* To whom correspondence should be addressed

Received January 25, 1993. Accepted March 5, 1993.

Calf thymus terminal deoxynucleotidyl transferase was used to incorporate several products of oxidative base damage onto the 3' end of oilgodeoxyribonucleotides. Under the defined conditions described in this report, single residues of dihydrothymine, ß-ureidoisobutyric acid, thymine glycol, urea, 7-hydro-8-oxoadenine, 7-hydro-8-oxoguanine, 5-hydroxycytosine and 5-hydroxyuracil were incorporated into oligodeoxyribonucleotides of different lengths. The reaction is both efficient and cost effective. The 3' termini of the reaction products were suitable substrates for ligation by phage T4 DNA ligase, facilitating greatly the construction of oligodeoxyribonucleotides containing unique and site specific oxidative DNA lesions.


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