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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:
- 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)
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