NDLI: A 32P-postlabeling method for detecting unstable N-7-substituted deoxyguanosine adducts in DNA.

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A 32P-postlabeling method for detecting unstable N-7-substituted deoxyguanosine adducts in DNA.

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A 32P-postlabeling method for detecting unstable N-7-substituted deoxyguanosine adducts in DNA.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Niu, T. Q. Austin-ritchie, P. Ludlum, D. B. Yu, D.
Description	Author Affiliation: Yu D ( Department of Pharmacology, University of Massachusetts Medical School, Worcester 01655.);
Abstract	Many antitumor agents, including the mustards, form N-7 deoxyguanosine adducts in DNA that are difficult to quantitate by the 32P-postlabeling procedure because of their instability. We have developed a method that is successful for the analysis of such adducts using, as a prototype mustard, 14C-labeled bis(2-chloroethyl)sulfide. This agent forms the unstable product 7-hydroxyethylthioethyldeoxyguanosine in DNA. By performing enzymatic digestions to 3'-deoxynucleotides at 10 degrees C, including a second N-7-substituted guanine deoxynucleotide as an internal standard, removing most of the unmodified nucleotides and [32P]ATP on disposable anion columns, and measuring the labeled products after separation on a C18 column, we are able to detect 1 unstable N-7 deoxyguanosine adduct in 10(7) normal nucleotides with good precision.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	15
Volume Number	91
Language	English
Publisher	National Academy of Sciences
Publisher Date	1994-08-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA Chemistry Deoxyguanosine Animals Chromatography, High Pressure Liquid DNA Damage Genetic Techniques Phosphorus Radioisotopes Research Support, U.S. Gov't, Non-P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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