Smolke:Protocols/Yeast Media Recipes

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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: 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)