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Nucleic Acids Research, 1980, Vol. 8, No. 2 377-387
© 1980

Article

Synchronization of HeLa cell cultures by inhibition of DNA polymerase a with aphidicolin

G. Pedrali-Noy*, S. Spadari*, A. Miller-Faurès+, A.O.A. Miller+, J. Kruppa{dagger} and G. Koch{dagger}

*Laboratorio di Genetica Biochimica ed Evoluzionistica, CNR, Pavia, Italy +Laboratory of Molecular Biochemistry, Faculty of Medicine, Mons State University, 7000 Mons, Belgium {dagger}Physiologisch-Chemisches Institut, Universität Hamburg, Abt. Molekularbiologie,Hamburg, GFR

Received December 1, 1979. Both the inhibitory effect of aphidicolin on the replicative {alpha}-polymerase and the reversibility of its action in vivo (Pedrali-Noy & Spadari, 1979, Biochem. Biophys. Res. Commun. 88, 1194–2002) allow the synchronization of cells in culture. Aphidicolin prevents G1 cells from entering the DNA synthetic period, blocks cells in "S" phase, allows G2, M and G1 cells to continue the cell cycle and to accumulate at the G1/S border. Aphidicolin is a more useful reagent than hydroxyurea and thymidine because it does not affect cell viability or "S" phase duration and does not interfere with the synthesis of dNTPs or DNA polymerases. In fact cells exposed to the drug continue to synthesize all three DNA polymerases {alpha}, ß and {gamma} as well as all dNTPs which, when the block is removed, are present at levels optimal for DNA initation and replication. The technique is simple amd can be applied to cells growing in suspension or monolayers and allows one to harvest large quantities of synchronized cells.


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