Aug 23, 2024

Public workspaceSolid Growth Medium - Yeast

DOI
dx.doi.org/10.17504/protocols.io.j8nlk85x5l5r/v1
  • 1UMass Chan Medical School, RNA Therapeutics Institute, Worcester, MA, USA;
  • 2University of Alberta, Department of Cell Biology, Edmonton, AB, Canada;
  • 3University of California, Department of Viticulture and Enology, Davis, CA, USA
Mathias Hammer
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DOI: dx.doi.org/10.17504/protocols.io.j8nlk85x5l5r/v1
Protocol Citation: Mathias Hammer, Ammeret Rossouw, Azra Lari, Ben Montpetit, David Grunwald 2024. Solid Growth Medium - Yeast. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk85x5l5r/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: April 05, 2024
Last Modified: August 23, 2024
Protocol Integer ID: 97849
Keywords: yeast grow medium, yeast solid grow medium
Funders Acknowledgement:
NSF
Grant ID: 1917206
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Abstract
This protocol describes the steps to prepare solid culture medium for Saccharomyces cerevisiae.
Materials
Chemicals:

SC-Ura Powder
Sunrise Science Products
Cat#: 1306-030
Lot#: 23K3083
Exp: 10/2027

Yeast Nitrogen Base Without Amino Acids
Sigma Life Science
Cat#: Y0626-250G
Lot#: SLBG0555V

Glucose
Sunrise Science Products
Cat#: 1907-1kg
Lot#: 3A0036

Agar
Sunrise Science Products
Cat#: 1910-500
Lot#: 3B0104

Sodium Hydroxide
Fisher Scientific
Cat#: S318-500
Lot#: 130802

Deionized Water

Equipment:

500 ml laboratory bottle with screw cap
1ml pipette
50 ml pipette
stirring hot plate
magnetic stirring bar
micro scales
autoclave
thermometer
10cm polystyrene Petri dishes (20 pieces)
4°C fridge
Before start
Have the following solutions premixed:

Glucose 20% 500 ml solution:
Concentration: 200 g/l
mix 100 g Glucose in 500 ml deionized water (ddH2O)

Sodium Hydroxide 1M solution 50 ml:
Concentration: 39.997 g/l
mix 1.99985 g NaOH into 50 ml ddH2O

Optional:
SC-xx 10x 100ml solution:
Concentration: 19.2 g/l
mix 1.92 g into 100 ml ddH2O

YNB 20x 100ml solution:
Concentration: 134.4 g/l
mix 13.44 g into 100 ml ddH2O
Compound medium for autoclave
Step case

Medium preparation with pre-resolved components
12 steps

This version of the protocol shows the preparation of the medium from SC-XX 10x and YNB 20x solutions.
Fill a 500 ml flask with Amount374 mL ddH2O.
Add a magnetic stirring bar and place the flask on a stirring hot plate.
Add Amount25 mL YNB 20x solution (Yeast Nitrogen Base with Ammonium Sulfate without Amino Acids).

Add Amount50 mL SC-XX 10x solution.
Note
In regard to cover all optional dropout media the amino acid base holds the notification -xx, where xx stand for the amino acid(s) that is as selection factor, missing in the medium.

Add Amount10 g Agar.
Add Amount1 mL NaOH 1 molar solution.
Note
This is essential for the solidification of the medium!

Autoclave for Duration00:15:00 at Temperature121 °C .
Note
Remove the stirring bar before going to autoclave.

Cooling and platting
Add a sterile stirring bar and thermometer. Let the medium stir to homogenize its temperature, while cooling.
When the medium cooled down to around Temperature80 °C add Amount50 mL sterile Glucose 20%.
When the medium cooled down to Temperature60 °C , plate Amount25 mL per Amount10 cm Petri dish (~20 dishes).
Let the medium solidify and cool down to TemperatureRoom temperature .
Seal the prepared dishes in a plastic bag to prevent condensation and store them in Temperature4 °C fridge.
Note
The agar plates can be store in the 4°C fridge for 2 to 3 months.