YPD media#

MATERIALS#

Reagents#

  • Bacto agar (if preparing agar plates)

  • Bacto peptone

  • Glucose (20%, w/v)

Sterilize by filtration or autoclaving.

  • Yeast extract (Fisher Scientific)

Equipment#

  • Autoclave

  • Flasks, autoclavable

  • Petri dishes, plastic, 9-cm diameter (if preparing agar plates)

  • Stir bar

  • Stir plate, magnetic

METHOD#

  1. To an autoclavable flask, add:

Reagent Amount to add (per 100mL)

Liquid

Agar plates

Bacto agar

-

2g

Bacto peptone

2g

2g

Yeast extract

1g

1g

Water

90mL

90mL

  1. Autoclave the mixture. (only if doing agar plates)

  2. Prepare as appropriate:

For liquid media

i. Add 10mL of sterile 20% (w/v) glucose. Mix.

ii. Allow to cool before use.

For agar plates

iii. Add 10mL of sterile 20% glucose per 100mL of media.

iv. While still warm, pour media (around 20ml) into 9-cm diameter plastic Petri dishes.

v. Allow agar mixture to cool and solidify.

SC – URA (20 ml SC-URA each)#

  • total amount: 400 mL

Protocol:

  • Add 360 mL of MiliQ

  • Add 2,72 g of yeast nitrogen base.

  • Add 0.308g of amino acid URA drop-out mix

  • Add 8g of Agar

  • Autoclave

  • Add 40 mL of 20 % dextrose (filter sterilized, add after autoclaving)

SC – LEU2 (20 ml SC-URA each)#

  • total amount: 400 mL

Protocol:

  • Add 360 mL of MiliQ

  • Add 2,72 g of yeast nitrogen base.

  • Add 0.276 g of amino acid LEU2 drop-out mix

  • Add 8g of Agar

  • Autoclave

  • Add 40 mL of 20 % dextrose (filter sterilized, add after autoclaving)

Special media:#

Low Fluorescence Complete Synthetic Media CSM-LF#

  • total amount: 400 mL

  • sugar source: Raffinose 20%

  • sugar final concentration: Raffinose 2%

Protocol:

  • Add 360 mL of MiliQ

  • Add 2,76 g of yeast nitrogen base. (005 bottle)

  • 0.316g CSM (009 bottle )

  • Filter stirilize

  • Add 40 mL of 20 % raffinose (filter sterilized, add after autoclaving)

YPD+6xADE+Agar#

  • total amount: 400 mL

  • sugar source: Dextrose 20%

  • sugar final concentration: Dextrose 2%

Protocol:

  • Add 60 mL of MiliQ

  • Add 4g of yeast extract

  • Add 8g peptone

  • Add 0.022g ADE

  • Filter stirilize

  • Autoclave 300ml MiliQ + 8g Agar

  • Poor the filter stirilize mixture to the liquid agar.

  • Add 40 mL of 20 % Dextrose (filter sterilized, add after autoclaving)

  • Poor 20ml to each plate.