Jan 06, 2023
Yeast Peptone Dextrose (YPD) medium
- 1University of Oslo
Protocol Citation: Andreas Sagen 2023. Yeast Peptone Dextrose (YPD) medium. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9d7x1g3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 05, 2023
Last Modified: January 06, 2023
Protocol Integer ID: 74794
Keywords: YPD, Yeasts, Saccharomyces cerevisiae, S. cerevisiae, yeast peptone dextrose, medium yeast peptone, yeast extract, yeast, others saccharomyces cerevisiae, many common yeast, glucose, peptone, protein, addition of protein, primary carbon
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.
Guidelines
Follow step by step, unless stated otherwise. Equipment needed should be standard to a microbiology lab.
Materials
Analytical scale, autoclave, bottle(s), weight vessel, LAF bench
Protocol materials
DextroseMerck MilliporeSigma (Sigma-Aldrich)Catalog #G7021
Yeast ExtractMerck MilliporeSigma (Sigma-Aldrich)Catalog #Y0875
PeptoneMerck MilliporeSigma (Sigma-Aldrich)Catalog #P5905
AgarMerck MilliporeSigma (Sigma-Aldrich)Catalog #A1296
Safety warnings
You can mix Dextrose from the beginning with the other compounds, and autoclave together. While this is more time efficient and easier, it is important to take into account the possibility of toxic byproducts produced by the Millard reaction when autoclaving, producing Acrylamide, a probable human carcinogen (IARC Group 2A).
Furthermore, when removing autoclaved components, be sure to take care as these can be very hot. If using antibiotics, use sufficient PPE to protect yourself, as some can be toxic to humans.
Before start
Prepare glassware by cleaning it, and ensure that scale is sufficiently calibrated
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 60 mL double-distilled water
Measure 10000 mg Dextrose
Powdered compounds:
DextroseMerck MilliporeSigma (Sigma-Aldrich)Catalog #G7021
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 00:05:00
5m
Adjust pH while mixing to 5.8 using concentrated sodium hydroxide
Add distilled water to a total of 100 mL
Autoclave liquid at 121 °C for 00:15:00
15m
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 300 mL double-distilled water
Measure 10000 mg Peptone and 5000 mg Yeast extract
Powdered compounds:
PeptoneMerck MilliporeSigma (Sigma-Aldrich)Catalog #P5905
Yeast ExtractMerck MilliporeSigma (Sigma-Aldrich)Catalog #Y0875
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 00:05:00
5m
Adjust pH while mixing to 5.8 using concentrated sodium hydroxide
Autoclave liquid at 121 °C for 00:15:00
15m
In a LAF bench, add 100 mL sterile Dextrose solution
Add sterile water to a total of 500 mL
Note
Cool to 50°C and supplement with antibiotics as appropriate
500 mL YPD (agar) medium
15m
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 300 mL double-distilled water
Measure 10000 mg Peptone, 5000 mg Yeast extract and 7500 mg agar
Powdered compounds:
PeptoneMerck MilliporeSigma (Sigma-Aldrich)Catalog #P5905
Yeast ExtractMerck MilliporeSigma (Sigma-Aldrich)Catalog #Y0875
AgarMerck MilliporeSigma (Sigma-Aldrich)Catalog #A1296
Add powdered solids into bottle, and use a magnetic mixer with a stir bar to mix for 00:05:00
5m
Adjust pH while mixing to 5.8 using concentrated sodium hydroxide
Autoclave liquid at 121 °C for 00:15:00
15m
In a LAF bench, add 100 mL sterile Dextrose solution
Add sterile water to a total of 500 mL
Note
Cool to 50°C and supplement with antibiotics as appropriate
Agar can be stored, then reheated to 50°C to be poured