Protocol

G₁/S Phase Synchronization Using Mimosine Arrest

This protocol was adapted from "Cell Synchronization," Chapter 14, in Cells (eds. Spector et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1998. This three-volume set is now out of print; however, some of the microscopy methods were republished in Basic Methods in Microscopy, by David L. Spector and Robert D. Goldman.

INTRODUCTION

This protocol uses the plant amino acid mimosine as a G₁/S synchronizing agent. Cells are first treated with excess thymidine to accumulate the majority of them at G₁/S; however, some cells will have stopped growth within the S phase. Thymidine is then removed to allow all the cells to proceed completely through the S phase. Mimosine is then added to arrest the cells at the G₁/S border. When mimosine is removed, cells will begin to enter S phase within about 1 hour.