Smolke:Protocols/Yeast Media Recipes

Yeast Nitrogen Base Minus Amino Acids (YNB-AA) Recipe
Autoclave (store at RT for media stocks)
 * 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

To use as working media:
 * After autoclaving add:
 * 100mL 10X (20%) Dextrose
 * 100mL 10X Drop Out stock (10X DO stock recipe here: [])
 * 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
Autoclave (store at RT for media stocks) To use as working media: After autoclaving add:
 * 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
 * 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 []

For yeast KanMX (G418) selection:
 * Must be in YP media (doesn't work with YNB)
 * 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)