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Use of an automated capillary DNA sequencer to investigate the interaction of cisplatin with telomeric DNA sequences
Author(s) -
Paul Moumita,
Murray Vincent
Publication year - 2012
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1664
Subject(s) - chemistry , dna , dna sequencer , cisplatin , capillary electrophoresis , adduct , dna sequencing , dna adduct , fluorescence , sequence (biology) , chromatography , biochemistry , genetics , biology , physics , organic chemistry , chemotherapy , quantum mechanics
The determination of the sequence selectivity of DNA‐damaging agents is very important in elucidating the mechanism of action of anti‐tumour drugs. The development of automated capillary DNA sequencers with fluorescent labelling has enabled a more precise method for DNA sequence specificity analysis. In this work we utilized the ABI 3730 capillary sequencer with laser‐induced fluorescence to examine the sequence selectivity of cisplatin with purified DNA sequences. The use of this automated machine enabled a higher degree of precision of both position and intensity of cisplatin–DNA adducts than previously possible with manual and automated slab gel procedures. A problem with artefact bands was overcome by ethanol precipitation. It was found that cisplatin strongly formed adducts with telomeric DNA sequences. Copyright © 2011 John Wiley & Sons, Ltd.