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RESEARCH DIVISION Laboratory Manual

 
 

Yeast Media

YEPD (non-selection)

- 1% yeast extract
- 2% peptone
- 1.5% agar (if needed for plates)

After autoclaving add glucose to 2% by adding 100 ml of 20% solution to each litre media.

SYNTHETIC MEDIA (selective media)

- 0.67% yeast nitrogen base without amino acids (Difco) 0919 15 3
- 1.5% agar (if needed for plates). Also add 1 pellet of NaOH per litre of agar to avoid mushy plates.

After autoclaving add 100 ml of 20% glucose (or galactose) solution and 100 ml of 10X Dropout solution to each litre media.

10X DROPOUT SOLUTION
- L-Isoleucine 300 mg/L
- L-Valine 1500mg/L
- L-Adenine hemisulfate salt 200 mg/L
- L-Arginine HCl 200 mg/L
- L-Lysine HCl 300 mg/L
- L-Methionine 200 mg/L
- L-Phenylalanine 500 mg/L
- L-Threonine 2000mg/L
- L-Tyrosine 300 mg/L
- L-Uracil 200 mg/L
- L-Histidine HCl monohydrate 200 mg/L* Sigma H9511
- L-Leucine 1000mg/L* Sigma L1512
- L-Tryptophan 200 mg/L* Sigma T0271

*    The 10X Dropout solution does not contain Histidine, Leucine and Tryptophan. These components are used to make selective media to select for yeast transformants.

** A better option is to purchase selective dropout amino acid mix. CSM -His,-Leu,-Trp -- BIO 101 (Cat. 10427-100) and add either His, Leu or Trp as needed.
Disclaimer: The following set of protocols were contributed by various members of our lab (past and present): Christine Andrews, Fiona Christensen, Neil Della, Ross Dickins, Debbie Donald, Andrew Holloway, Gary Hime, Colin House, Yinling Hu, Rachael Parkinson, Nadia Traficante, Hannah Robertson, Ping Fu and Dennis Wang. Special thanks to Vicki Hammond, Frank Kontgen and Maria Murphy, who contributed many of the ES cell protocols. Sections dealing with Photomicroscopy, Polyclonal and Monoclonal Antibody Production were provided by members of Gerry Rubin's Laboratory (Berkeley). Any comments in the methods (technical errors etc.) E-mail: d.bowtell@pmci.unimelb.edu.au
David Bowtell PMCI October 1998