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. 1991 Jun 11;19(11):2929–2933. doi: 10.1093/nar/19.11.2929

Measurement of the sequence specificity of covalent DNA modification by antineoplastic agents using Taq DNA polymerase.

M Ponti 1, S M Forrow 1, R L Souhami 1, M D'Incalci 1, J A Hartley 1
PMCID: PMC328253  PMID: 2057351

Abstract

A polymerase stop assay has been developed to determine the DNA nucleotide sequence specificity of covalent modification by antineoplastic agents using the thermostable DNA polymerase from Thermus aquaticus and synthetic labelled primers. The products of linear amplification are run on sequencing gels to reveal the sites of covalent drug binding. The method has been studied in detail for a number of agents including nitrogen mustards, platinum analogues and mitomycin C, and the sequence specificities obtained accord with those obtained by other procedures. The assay is advantageous in that it is not limited to a single type of DNA lesion (as in the piperidine cleavage assay for guanine-N7 alkylation), does not require a strand breakage step, and is more sensitive than other primer extension procedures which have only one cycle of polymerization. In particular the method has considerable potential for examining the sequence selectivity of damage and repair in single copy gene sequences in genomic DNA from cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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