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Matsumoto Yoshihiro
Base excision repair in mammalian cells
Methods in Molecular Biology (Totowa, NJ, United States). 2006 ;314(DNA Repair Protocols (2nd Edition)) :365-375
PMID: AN 2005:1186502   
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Abstract
A rapid, convenient and safe in vitro assay system for base excision repair is described. Whole cell exts. are prepd. by detergent-based cell lysis and provide a vigorous activity of AP site repair. A circular DNA substrate is used for detection of both DNA polymerase b-dependent and proliferating cell nuclear antigen (PCNA)-dependent pathways. Repaired and unrepaired DNA substrates are sepd. by agarose gel electrophoresis as a linear DNA mol. and a nicked circular mol., resp., and detected by staining with SYBR Green I. This assay system does not require radioactive substrates or nucleotides, and provides a sensitivity in which 10 ng of a DNA substrate per reaction is sufficient for quant. repair anal. [on SciFinder (R)]
Notes
3 Biochemical Genetics Department of Pharmacology,Fox Chase Cancer Center,Philadelphia,PA,USA. Journal 1064-3745 written in English.