Growth and Manipulation of Yeast

Douglas A. Treco1, Ann Reynolds2, Victoria Lundblad3

1 Massachusetts General Hospital, Boston, Massachusetts, 2 University of Washington, Seattle, Washington, 3 Baylor College of Medicine, Houston, Texas
Publication Name:  Current Protocols in Protein Science
Unit Number:  Appendix 4L
DOI:  10.1002/0471140864.psa04ls14
Online Posting Date:  May, 2001
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Abstract

This unit describes preparation of selected media for growing yeast and also discusses strain storage and revival. Protocols are provided for the assay of β‐galactosidase in liquid culture and for transformation using lithium acetate.

     
 
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Table of Contents

  • Preparation of Selected Yeast Media
  • Strain Storage and Revival
  • Basic Protocol 1: Assay for β‐Galactosidase in Liquid Cultures
  • Basic Protocol 2: Transformation Using Lithium Acetate
  • Reagents and Solutions
  • Literature Cited
  • Tables
     
 
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Materials

Basic Protocol 1: Assay for β‐Galactosidase in Liquid Cultures

  Materials
  • YPD medium (see recipe under Liquid Media)
  • recipeZ buffer (see recipe in Reagents and Solutions)
  • 0.1% sodium dodecyl sulfate(SDS)
  • Chloroform
  • 4 mg/ml O‐nitrophenyl‐β‐D‐galactoside (ONPG) in 0.1 M potassium phosphate, pH 7.0 (see appendix 2E for buffer; filter sterilize and store frozen)
  • 1 M Na 2CO 3
  • 30°C water bath

Basic Protocol 2: Transformation Using Lithium Acetate

  Materials
  • YPD medium (see recipe under Liquid Media)
  • Yeast strain to be transformed
  • YPAD medium: YPD medium supplemented with 30 mg/liter adenine hemisulfate
  • Highest‐quality sterile H 2O
  • 10× TE buffer, pH 7.5 (see appendix 2E for 1× recipe), sterile
  • 10× lithium acetate stock solution: 1 M lithium acetate, pH 7.5 (adjust pH with dilute acetic acid), filter sterilized
  • DNA: high‐molecular‐weight, single‐stranded carrier DNA and transforming DNA
  • 50% (w/v) polyethylene glycol (PEG 4000 or 3350 (do not use PEG 8000), filter sterilized
  • CM dropout plates (see recipe under Solid Media) prepared with Difco agar
  • 30°C incubator with shaker
  • Sorvall GSA and SS‐34 rotors (or equivalents)
  • 42°C water bath
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Figures

Videos

Literature Cited

Literature Cited
   Sherman, F., Fink, G.R., and Lawrence, C.W. 1979. Methods in Yeast Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
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