Smolke:Protocols/Yeast Media Recipes

Yeast Nitrogen Base Minus Amino Acids (YNB-AA) Recipe

• For 1 L Batch:
  • 1.7g Yeast Nitrogen Base (stored in cold room)
  • 5g NH3SO4 (Ammonium Sulfate, stored in chemical stock cabinet)
• Dissolve in 800mL Np H2O for YNB-AA stock
• Adjust pH value to 5.8 using NaOH (starting May 2011) -Yen-Hsiang

Autoclave (store at RT for media stocks)

To use as working media:

• After autoclaving add:
  • 100mL 10X (20%) Dextrose
  • 100mL 10X Drop Out stock (10X DO stock recipe here: [[1]])
  • 1 L Total Volume

Yeast Nitrogen Base Minus Amino Acids Non-Inducing, Non-Repressing (YNB-AA NINR) Recipe

• For 1 L Batch:
  • 1.7g Yeast Nitrogen Base
  • 5g NH3SO4
• Dissolve in 700mL Np H2O for YNB-AA NINR stock
• Adjust pH value to 5.8 using NaOH (starting May 2011) -Yen-Hsiang
• Autoclave (store at RT for media stocks)

To use as working media:

• After autoclaving add:
  • 100mL 10X (20%) Raffinose
  • 100mL 10X (10%) Sucrose
  • 100mL 10X Drop Out stock
  • 1 L Total Volume

Yeast Peptone (YP) Media Recipe

• For 1 L Batch:
  • 10g Yeast Extract [Bacto™ REF#212750] (stored in chemical stock cabinet)
  • 20g Peptone [Bacto™ REF#211677] (stored in chemical stock cabinet)
  • 80 mg of Adenine hemisulfate (Sigma Chemical Co. Ltd., cat. no. A-3159) (stored in chemical stock cabinet)
  • Dissolve in 900mL Np H2O for YP stock

Autoclave (store at RT for media stocks) To use as working media: After autoclaving add:

• • 100mL 10X (20%) Dextrose
  • 1 L Total Volume YPD (Yeast Peptone Dextrose)

To pour any of these media stocks as plates:

• • Add 20g agar per 1L final media volume before autoclaving
  • See here [[2]]

For yeast KanMX (G418) selection:

• Cannot use ammonium sulfate as primary nitrogen source
• Add G418 to YP or to YNB with 1 g/l MSG instead of ammounium sulfate
• Frozen stock is good for 3 months.
• dry G418 is stored in chemical room.
• Add 400mg/L for selection (in plates)
• Add 200mg/L for maintenance (in liquid culture media)

-Michael S. Siddiqui 21:53, 4 January 2011 (EST)* 01/04/2011 (based mainly on communications from Andrew Babiskin)