Growth and Manipulation of Yeast

Douglas A. Treco1, Fred Winston2

1 Massachusetts General Hospital, Boston, Massachusetts, 2 Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 13.2
DOI:  10.1002/0471142727.mb1302s82
Online Posting Date:  April, 2008
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Yeast cultures can be grown, maintained, and stored in liquid media or on agar plates using techniques similar to those for bacterial cultures. This unit describes culture conditions for these basic techniques. Additional methods describe determination of yeast mating type, diploid construction, sporulation, tetrad dissection, and random spore analysis. Curr. Protoc. Mol. Biol. 82:13.2.1‐13.2.12. © 2008 by John Wiley & Sons, Inc.

Keywords: yeast; media; yeast culture; growth

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Basic Protocol 1: Growth in Liquid Media
  • Basic Protocol 2: Growth on Solid Media
  • Basic Protocol 3: Determination of Cell Density
  • Basic Protocol 4: Determination of Phenotype by Replica Plating
  • Basic Protocol 5: Determination of Mating Type
  • Basic Protocol 6: Diploid Construction
  • Basic Protocol 7: Sporulation of Diploid Cells
  • Basic Protocol 8: Preparation and Dissection of Tetrads
  • Support Protocol 1: Preparation of Dissecting Needles
  • Alternate Protocol 1: Random Spore Analysis
  • Commentary
  • Figures
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Growth in Liquid Media

  Materials
  • S. cerevisiae: MATα thr4 (tester), MATα thr4 (tester), and uncharacterized strains
  • YPD medium and plates (unit 13.1)
  • Minimal plates (unit 13.1)
  • Replica plating block (unit 1.3)
  • Sterile velvets (unit 1.3)

Basic Protocol 2: Growth on Solid Media

  Materials
  • Yeast cells
  • Sporulation plates or sporulation medium, with appropriate nutrients (unit 13.1)
  • YPD medium (unit 13.1)

Basic Protocol 3: Determination of Cell Density

  Materials
  • Spores (see protocol 7)
  • 0.5 mg/ml Zymolyase‐100T (ICN Immunobiologicals) in 1 M sorbitol
  • Dissecting microscope
  • Dissecting needle (see Support Protocol)

Basic Protocol 4: Determination of Phenotype by Replica Plating

  Materials
  • Spores (see protocol 7)
  • 1 mg/ml Zymolyase‐20T (ICN Immunobiologicals) in H 2O, filter sterilized
  • 2‐Mercaptoethanol (2‐ME)
  • 1.5% (v/v) Nonidet P‐40 (NP‐40)
  • Ethanol
  • Sonicator and probe
  • Additional reagents and equipment for cell counting (unit 1.2) and replica plating (unit 1.3)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Esposito, R.E. and Klapholz, S. 1981. Meiosis and ascosporal development. In The Molecular Biology of Yeast Saccharomyces: Life Cycle and Inheritance ( J.N. Strathern, E.W. Jones, and J.R. Broach, eds.) pp. 211‐287. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
   Sherman, F. 2002. Getting started with yeast. In Guide to Yeast Genetics and Molecular and Cell Biology, Part B (C. Guthrie and G.R. Fink, eds.) pp. 3‐41. Academic Press, San Diego.
   Wang, H‐T., Frackman, S., Kowalisyn, J., Esposito, R.E., and Elder, R. 1987. Developmental regulation of SPO13, a gene required for separation of homologous chromosomes at meiosis I. Mol. Cell. Biol. 7: 1425‐1435.
Key Reference
   Sherman, F., Fink, G.R., and Lawrence, C.W. 1979. Methods in Yeast Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
  Provides a number of detailed procedures for genetic experiments that may be of interest to more advanced students.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library