Yeast Peptone Dextrose (YPD) medium
Andreas Sagen
Abstract
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
Steps
100 mL Dextrose solution
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
Fill the bottle with 60mL
double-distilled water
Measure 10000mg
Dextrose
Powdered compounds:
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 0h 5m 0s
Adjust pH while mixing to 5.8
using concentrated sodium hydroxide
Add distilled water to a total of 100mL
Autoclave liquid at 121°C
for 0h 15m 0s
500 mL YPD (broth) medium
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
Fill the bottle with 300mL
double-distilled water
Measure 10000mg
Peptone and 5000mg
Yeast extract
Powdered compounds:
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 0h 5m 0s
Adjust pH while mixing to 5.8
using concentrated sodium hydroxide
Autoclave liquid at 121°C
for 0h 15m 0s
In a LAF bench, add 100mL
sterile Dextrose solution
Add sterile water to a total of 500mL
500 mL YPD (agar) medium
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
Fill the bottle with 300mL
double-distilled water
Measure 10000mg
Peptone, 5000mg
Yeast extract and 7500mg
agar
Powdered compounds:
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 0h 5m 0s
Adjust pH while mixing to 5.8
using concentrated sodium hydroxide
Autoclave liquid at 121°C
for 0h 15m 0s
In a LAF bench, add 100mL
sterile Dextrose solution
Add sterile water to a total of 500mL