Protocols (Materials and Methods)
Media Formulations

 

Index:
(to jump to a listing, click on the desired name to the right)

MCDB (Modified Czapek Dox) Broth
Corn Meal Agar

CDN Agar
CDY (Czapek Dox Yeast Extract)
Malt Extract/Peptone (ME/P)
Minimal Medium (SD)
YPD Broth
 

  Formulations: Media recipes
  Media Recipes

MCDB (Modified Czapek Dox) Broth

(Oujezdsky K.B., Grove S.N., and P.J. Szaniszlo. 1972. Morphological and structural changes during the yeast-to-mold conversion of Phialophora dermatitidis. J. Bacteriol. 113:468-477.)

Czapek Dox broth (Difco) w/ 1.0% yeast extract (Difco)

This medium is made using pre-prepared Difco Czapek Dox broth which contains sucrose as a carbon-source.

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Corn Meal Agar

 

17g Corn Meal Agar (Difco)

10g agar (Difco)

 

CDN Agar

Into 800 ml distilled H20 add:
10 ml 30% NaNO3
5 ml 20% K2HPO4
5 ml 20% MgSO4:7H2O
5 ml 10% KCl
30 mg glucose/dextrose

Bring to 1 liter and adjust pH to 6.5, then add:

3 mg thiamine / L
5.3g NH4Cl / L

If necessary readjust to pH of 6.5, then add 20g Bacto-agar

CDN is CDY broth in which the yeast extract component was replaced with 3 mg of thiamine and 5.3 g of NH4Cl for each liter of medium. For determination of auxotrophy, CDN was supplemented with adenine, arginine, methionine, and uracil as appropriate.

Solid media were prepared by adding 2% (wt/vol) Bacto agar (Difco) or 1% (wt/vol) GelRIte (Kelco, San Diego, Calif.) prior to autoclaving.

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CDY (Czapek Dox Yeast Extract)

Add to 800 ml distilled water

10 ml 30% NaNO3
5 ml 20% K2HPO4
5 ml 20% MgSO4:7H2O
5 ml 10% KCl
30 mg glucose/dextrose

Adjust pH to 6.5

Add 1 g Bacto yeast extract
1 ml fresh FeSO47H2O (10ml /10ml)

adjust to 1 L by H20

For solid: add 20 g of agar

CDY (g/l)

Adapted from (Roberts, R.L., and P.J. Szaniszlo. 1978. Temperature-sensitive multicellular mutants of Wangiella dermatitidis. J. Bacteriol. 135:622-632.)

30 g/l dextrose,
3 g/l NaNO
3
1 g/l K2HPO4
0.5 g/l MgSO4

Adjust to pH of 6.5

0.01 g/l FeSO4

Adjust to pH of 6.5

1 g/l yeast extract (Difco)

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Malt Extract/Peptone (ME/P)

30 g malt extract
10 g peptone
1 L dd-H20

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Minimal Medium (SD)

1.7 gm yeast nitrogen base
without amino acids and } Autoclave
without ammonium sulfate

5 gm ammonium sulfate

400 ml H20
20 gm Bacto-agar
500 ml H20 } Autoclave

After autoclave, mix 2 together and add 50 ml 40% glucose

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YPD Broth

10g Yeast Extract
20 g Poly peptone
20 g Dextrose
1L H20

mix and autoclave

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