TITLE: Yeast Peptone Dextrose (YPD) medium
AUTHORS: Andreas Sagen

[DESCRIPTION]
Yeast peptone dextrose is a medium composition used to grow many common yeasts, among others Saccharomyces cerevisiae. Glucose is the primary carbon and energy source, while nitrogen and essential amino acids are provided by the yeast extract and peptone. It is nonselective and the addition of protein and yeast extract allows faster growth so that during exponential or log-phase growth, the cells divide every 90 minutes in S. cerevisiae.

[BEFORE_START]
Prepare glassware by cleaning it, and ensure that scale is sufficiently calibrated

[GUIDELINES]
Follow step by step, unless stated otherwise. Equipment needed should be standard to a microbiology lab.

[STEPS]
SECTION: 100 mL Dextrose solution
1. All compounds are measured using a high precision analytical scale from powdered compounds. Each compound is measured to within 1% of the target weight. All compounds are mixed in a Duran bottle
SECTION: 500 mL YPD (broth) medium
2. All compounds are measured using a high precision analytical scale from powdered compounds. Each compound is measured to within 1% of the target weight. All compounds are mixed in a Duran bottle
SECTION: 500 mL YPD (agar) medium
3. All compounds are measured using a high precision analytical scale from powdered compounds. Each compound is measured to within 1% of the target weight. All compounds are mixed in a Duran bottle
SECTION: 100 mL Dextrose solution
1.1. Fill the bottle with 60 mL double-distilled water
SECTION: 100 mL Dextrose solution
1.4. Adjust pH while mixing to pH 5.8 using concentrated sodium hydroxide
SECTION: 100 mL Dextrose solution
1.3. Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 5 min

SECTION: 100 mL Dextrose solution
1.2. Measure 10000 mg Dextrose

Powdered compounds:
 
SECTION: 100 mL Dextrose solution
1.5. Add distilled water to a total of 100 mL
SECTION: 100 mL Dextrose solution
1.6. Autoclave liquid at 121 °C for 15 min
SECTION: 500 mL YPD (broth) medium
2.1. Fill the bottle with 300 mL double-distilled water
SECTION: 500 mL YPD (broth) medium
2.5. Autoclave liquid at 121 °C for 15 min
SECTION: 500 mL YPD (broth) medium
2.4. Adjust pH while mixing to pH 5.8 using concentrated sodium hydroxide
SECTION: 500 mL YPD (broth) medium
2.3. Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 5 min
SECTION: 500 mL YPD (broth) medium
2.2. Measure 10000 mg Peptone and 5000 mg Yeast extract

Powdered compounds:
 
 
SECTION: 500 mL YPD (broth) medium
2.6. In a LAF bench, add 100 mL sterile Dextrose solution
SECTION: 500 mL YPD (broth) medium
2.7. Add sterile water to a total of 500 mL
 

SECTION: 500 mL YPD (agar) medium
3.1. Fill the bottle with 300 mL double-distilled water
SECTION: 500 mL YPD (agar) medium
3.2. Measure 10000 mg Peptone, 5000 mg Yeast extract and 7500 mg agar

Powdered compounds:
 
 
 
SECTION: 500 mL YPD (agar) medium
3.3. Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 5 min

SECTION: 500 mL YPD (agar) medium
3.4. Adjust pH while mixing to pH 5.8 using concentrated sodium hydroxide
SECTION: 500 mL YPD (agar) medium
3.5. Autoclave liquid at 121 °C for 15 min
SECTION: 500 mL YPD (agar) medium
3.6. In a LAF bench, add 100 mL sterile Dextrose solution
SECTION: 500 mL YPD (agar) medium
3.7. Add sterile water to a total of 500 mL