By Daryl S. Henderson
Univ. of Dundee, united kingdom. offers specified directions for learning manifold features of the eukaryotic reaction to genomic harm. Emphasizes equipment for interpreting DNA fix approaches in mammalian cells. For researchers. Hardcover, plastic-comb binding additionally on hand.
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Additional resources for DNA Repair Protocols: Eukaryotic Systems
Evans, D. and Phillips, J. P. (1992) Urate-null mutants of Drosophila melanogaster are hypersensitive to oxygen stress. Proc. Natl. Acad. Sci. USA 89, 4343–4347. 34. Vogel, E. and Natarajan, A. T. (1979) The relation between reaction kinetics and mutagenic action of mono-functional alkylating agents in higher eukaryotic systems. I. Recessive lethal mutations and translocations in Drosophila. Mutat. Res. 62, 51–100. 30 Henderson 35. Ashburner, M. (1989) Drosophila: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Vortex to dissolve. 15. Methyl methanesulfonate (MMS) (Merck): Prepare as for EMS. (1% v/v; 118 mM). 16. Mitomycin C (MMC) (Kyowa Hakko, Kogyo Co. Ltd. Tokyo, Japan), stock solution (4 mg/mL): Dissolve in sterile H2O (or PBS). Keep at 0–4°C. 17. Bleomycin (BLM) (Lundbeck B. V. Amsterdam, Holland), freshly dissolved in PBS. Mutagen-Sensitive CHO Cell Lines 51 3. Methods Culture cells in 10- or 15-cm plastic dishes containing 10 or 25 mL of standard medium, respectively, at 37°C in an atmosphere of 5% CO2 in a humidified incubator.
3. Irradiate the seedlings with a dose of 200 J/m2 UV-C from an unfiltered germicidal lamp (see Note 6). To eliminate photoreactivation of UV-induced dimers, it is important to avoid exposure to blue or UV-A radiation after the UV treatment. Set up your germicidal lamp in a dark room with a red safety light. Allow the UV lamp to warm up for 15 min prior to use; its spectral qualities will change and then stabilize during this time (11). Caution: Protect your skin, especially your eyes and lips, from UV-C radiation by using a plastic face shield, lab coat, and gloves.
DNA Repair Protocols: Eukaryotic Systems by Daryl S. Henderson