Cell-Cycle Analysis of Saccharomyces cerevisiae

This chapter discusses flow cytometry measurement methods of the DNA content of Saccharomyces cerevisiae populations, although the analysis methods might be applicable to other yeasts with minor modifications. The chapter describes methods for staining yeast DNA with mithramycin, propidium iodide, and acriflavin. The bromodeoxyuridine (BrdUrd) method, as adapted for yeast, is also described. The chapter discusses the mathematical analysis of yeast DNA histograms. Typical DNA histograms from a diploid strain of S. cerevisiae stained with mithramycin, acriflavine, and propidium iodide are displayed and cell-cycle fractions estimated from these histograms using Multicycle are summarized. Representative data obtained with each of the protocols and the subsequent mathematical analyses are compared in the chapter. All data were obtained from the same sample from an exponentially growing yeast population. In addition, an example is given in which the BrdUrd labeling technique is applied 10 measure the distributions of cell properties at specific points within the cell cycle.