Unit

UNIT 11.13 Cell Cycle Analysis of Yeasts

  1. Margarida Fortuna1,
  2. Maria João Sousa1,
  3. Manuela Côrte-Real1,
  4. Cecília Leão1,
  5. Alexandre Salvador2,
  6. Filipe Sansonetty2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142956.cy1113s13

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Fortuna, M., João Sousa, M., Côrte-Real, M., Leão, C., Salvador, A. and Sansonetty, F. 2001. Cell Cycle Analysis of Yeasts. Current Protocols in Cytometry. 13:11.13:11.13.1–11.13.9.

Author Information

  1. 1

    Universidade do Minho, Braga, Portugal

  2. 2

    IPATIMUP, Porto, Portugal

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 2000

Abstract

Staining protocols generally designed for the flow cytometric analysis of the cell cycle in mammalian cells are frequently not satisfactory for quantification of the various cell-cycle phases in yeasts. High CVs limit the accuracy of DNA content measurement and estimates of populations in cell-cycle compartments. This unit describes a staining procedure for yeasts using the sensitive nucleic acid stain SYBR Green I, which binds to double-stranded DNA with high selectivity and which has a much higher fluorescence quantum yield upon binding than most other commonly used fluorophores. The properties of this dye combined with optimized sample processing provide high-resolution DNA analysis, with half-peak CVs around 3 to 4% and clear-cut identification of the S phase.