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.