Synchronization of cells.

Abstract

Synchronization of cells is essential to study cell cycle specific events. If, for example, one suspects that a given DNA repair pathway is used in a particular cell cycle phase, the protocol can be used to enrich cells in each phase of the cell cycle and analyze the cellular response to DNA damage. Synchronization is also useful, when a gene is essential for a particular phase of the cell cycle. If a gene is, for example, essential for mitosis, synchronization of the cells in G1 phase with concomitant inactivation of the gene enables us to study the function of the gene in interphase, and to follow synchronous cell cycle progression to M phase. Two synchronization methods: centrifugal elutriation to enrich G1, S or G2 phase cells and nocodazole-mimocine sequential treatment to enrich cells at the G1/S boundary are described. Centrifugal elutriation can be achieved in less time (0.5-2 h) and with very little physiological stress to the cells whereas synchronization by drugs, such as nocodazole and mimocine, may result in unfavorable side effects.