Single Cell Microarray for High Throughput Detection of DNA Damage

LEAD AUTHOR Bevin P. Engelward (MIT-Cambridge) Principal Investigator Biological Engineering MIT Cambridge, MA USA

OTHER AUTHORS: Jing Ge, Danielle Chow Jessica Fessler
PROTOCOL TYPE: Novel Technologies
Protocol Document
SYNOPSIS: Motivation • DNA damage is associated with an increased risk of cancer, aging and disease • Few methods are available for high throughput analysis of DNA damage • This protocol is based upon the traditional comet assay wherein DNA migrates more readily through a matrix when damaged • 96 samples can be processed in parallel • Throughput and sensitivity are increased compared to the traditional assay
PUBLICATIONS: J. Ge, D. K. Wood, D. M. Weingeist, S. Prasongtanakig, P. Navasumrit, M. Ruchirawat, B.P. Engelward. Standard fluorescent imaging of live cells is highly genotoxic,Cytometry A, 83:552-560 (2013). D. M. Weingeist, J. Ge, D. K. Wood, J. T. Mutamba, Q. Huang, E. A. Rowland, M. B. Yaffe, S. Floyd, and B. Engelward. Single cell microarray enables high throughput evaluation of DNA double strand breaks and DNA repair inhibitors, Cell Cycle, 12:907-915(2013). M. W. Chao, M. Y. Kim, W. Ye, J. Ge, L. J. Trudel, C. L. Belanger, P. L. Skipper, B. P. Engelward, S. R. Tannenbaum, G. N. Wogan. Genotoxicity of 2,6- and 3,5-dimethylaniline in cultured Mammalian cells: the role of reactive oxygen species. Toxicol Sci. 130(1):48-59 (2012). D. K. Wood, D. M. Weingeist, Y. Wu, S. N. Bhatia and B. P. Engelward, Single cell trapping and DNA damage an alysis using microwell arrays, Proc. Natl. Acad. Sci. USA,107:10008-10013 (2010).

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