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

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
• 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)

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 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 selection: 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)