Yeast Peptone Dextrose (YPD) medium

Andreas Sagen

Published: 2023-01-06 DOI: 10.17504/protocols.io.x54v9d7x1g3e/v1

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

1.1.

Fill the bottle with 60mL double-distilled water

1.2.

Measure 10000mg Dextrose

Powdered compounds:

1.3.

Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 0h 5m 0s

1.4.

Adjust pH while mixing to 5.8 using concentrated sodium hydroxide

1.5.

Add distilled water to a total of 100mL

1.6.

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

2.1.

Fill the bottle with 300mL double-distilled water

2.2.

Measure 10000mg Peptone and 5000mg Yeast extract

Powdered compounds:

2.3.

Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 0h 5m 0s

2.4.

Adjust pH while mixing to 5.8 using concentrated sodium hydroxide

2.5.

Autoclave liquid at 121°C for 0h 15m 0s

2.6.

In a LAF bench, add 100mL sterile Dextrose solution

2.7.

Add sterile water to a total of 500mL

Note

Cool to 50°C and supplement with antibiotics as appropriate

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

3.1.

Fill the bottle with 300mL double-distilled water

3.2.

Measure 10000mg Peptone, 5000mg Yeast extract and 7500mg agar

Powdered compounds:

3.3.

Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 0h 5m 0s

3.4.

Adjust pH while mixing to 5.8 using concentrated sodium hydroxide

3.5.

Autoclave liquid at 121°C for 0h 15m 0s

3.6.

In a LAF bench, add 100mL sterile Dextrose solution

3.7.

Add sterile water to a total of 500mL

Note

Cool to 50°C and supplement with antibiotics as appropriateAgar can be stored, then reheated to 50°C to be poured

Yeast Peptone Dextrose (YPD) medium

Abstract

Before start

Steps

100 mL Dextrose solution

500 mL YPD (broth) medium

500 mL YPD (agar) medium

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