Yeast Media



Hartwell's Complete (HC) solution

To make 4L of 10x HC (6 Dropout Amino Acid Liquid) stock:

  • 0.8 g ARG
  • 4.0 g ASP
  • 4.0 g GLU
  • 3.2 g ILE
  • 2.0 g PHE
  • 16.0 g SER
  • 8.0 g THR
  • 2.4 g TYR
  • 6.0 g VAL
Dissolve in a final volume of 4L and filter sterilize.

Yeast Complete (YC) solution

To make 1L of 100x YC (7 Dropout Amino Acid Liquid) stock:

  • 10.0 g ARG
  • 5.0 g ASP
  • 5.0 g ILE
  • 5.0 g PHE
  • 5.0 g PRO
  • 5.0 g SER
  • 10.0 g THR
  • 5.0 g VAL
Dissolve in a final volume of 1L and filter sterilize.

Yeast Nitrogen Base (YNB) Solution

To make 4L 10x YNB solution:

  • 68 g Yeast Nitrogen Base (w/o amino acids and w/o ammonium sulfate)
  • 200 g ammonium sulfate
  • (Optional) 2.0 g TYR
Dissolve in a final volume of 4L and autoclave.

Common Supplement Solutions

Table of Common Supplements
SUPPLEMENT STOCK SOL'N [STOCK] USE VOL
Adenine Hemisulfate 100 mg/100 ml 100X (1 mg/ml) 10 ml/L
Uracil 200 mg/100 ml 100X (2 mg/ml) 100 ml/L
L-Histidine 500 mg/ 100 ml 100X (5 mg/ml) 10 ml/L
L-Arginine 1 g/100 ml 100X (10 mg/ml) 10 ml/L
L-Methionine 500 mg/100 ml 100X (5 mg/ml) 10 ml/L
L-Leucine 1 g/ 100ml 100X (10 mg/ml) 10 ml/L
L-Lysine 1 g/ 100ml 100X (10 mg/ml) 10 ml/L
L-Tryptophan* 1 g/ 100ml 100X (10 mg/ml) 10 ml/L

*L-Tryptophan is sterilized through a sterile filter, not autoclaved, and is stored at 4oC.

Hartwell Complete (HC) Plates

To make 1L Hartwell Complete (HC) plates:

  • 20 g of Agar
  • 540 ml distilled H2O
Autoclave in a 2L flask for 20 minutes.

After autoclaving, add the following:
  • 100 ml 20% sterile glucose
  • 100 ml 10X YNB solution
  • 100 ml 10X HC-7aa dropout solution
  • 100 ml URA solution
  • 10 ml ADE solution
  • 10 ml LYS solution
  • 10 ml MET solution
  • 10 ml TRP solution
  • 10 ml LEU solution
  • 10 ml HIS solution

Yeast Complete (YC) Plates

To make 1L Yeast Complete (YC) plates:

  • 20 g of Agar
  • 630 ml distilled H2O
Autoclave in a 2L flask for 20 minutes.

After autoclaving, add the following:
  • 100 ml 20% sterile glucose
  • 100 ml 10X YNB solution
  • 10 ml 100X YC-7aa dropout solution
  • 100 ml URA solution
  • 10 ml ADE solution
  • 10 ml LYS solution
  • 10 ml MET solution
  • 10 ml TRP solution
  • 10 ml LEU solution
  • 10 ml HIS solution

FOA Plates

To make 1L YC plus FOA media:

  • 20 g of Agar
  • 630 ml distilled H2O
Autoclave in a 2L flask for 20 minutes.

After autoclaving, add the following:
  • 100 ml 20% sterile glucose
  • 100 ml 10X YNB solution
  • 10 ml 100X YC-8aa dropout solution
  • 100 ml URA solution
  • 10 ml ADE solution
  • 10 ml LYS solution
  • 10 ml MET solution
  • 10 ml TRP solution
  • 10 ml LEU solution
  • 10 ml HIS solution
Let flask cool to 50oC, then add 1.0 g of 5-FOA.


Casamino Acid (CAA) Plates

To make 1L Casamino Acid (CAA) media:

  • 20 g of Agar
  • 6.7 g Yeast Nitrogen Base without amino acids
  • 20 g Casamino acids
  • 860 ml distilled H2O
Agar must be washed three times with distilled H2O to rinse out trace amounts of uracil prior to preparation.

Autoclave in a 2L flask for 20 minutes.

After autoclaving, add the following:
  • 100 ml 20% sterile glucose
  • 10 ml 100X CAA supplement
  • 10 ml 100X TRP supplement

To make 1L 100X CAA Supplement:
  • 2 g ADE
  • 2 g HIS
  • 3 g LEU
  • 3 g LYS
  • 2 g MET
  • 500 ml distilled H2O
Sterilize through a sterile filter.

To make 200ml 100X TRP Supplement:
  • 0.4 g TRP
  • 200 ml distilled H2O
Sterilize through a sterile filter.


Yeast-extract Peptone Dextrose (YPD) Plates

To make 1L Yeast-extract Peptone Dextrose (YPD) media:

  • 20 g of Agar
  • 10 g Yeast Extract
  • 20 g Bacto-peptone
  • 860 ml distilled H2O
Autoclave in a 2L flask for 20 minutes.

After autoclaving, 100 ml 20% sterile glucose.

Really Rich Media (RRM) Plates

To make 1L Really Rich Media (RRM):

  • 20 g of Agar
  • 10 g Yeast Extract
  • 2.5 g Bacto-tryptone
  • 30 g Nutrient Broth powder
  • 680 ml distilled H2O
CAUTION: This media has a tendency to boil over, therefore a large flask is needed. Autoclave in a 4L flask for 20 minutes.

Exercise care when removing the flask from the autoclave.

After autoclaving, add 175 ml 20% sterile glucose.

Minimal Sporulation Plates

To make 1L Minimal Sporulation media:

  • 20 g of Agar
  • 10 g KOAc
  • 950 ml distilled H2O
Autoclave in a 2L flask for 20 minutes.