Smolke:Protocols/Yeast Media Recipes: Difference between revisions
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'''For yeast KanMX selection:''' | '''For yeast KanMX selection:''' | ||
**Must be in YP media (doesn't work with YNB) | |||
**Add 400mg/L for selection (in plates) | **Add 400mg/L for selection (in plates) | ||
**Add 200mg/L for maintenance (in liquid culture media) | **Add 200mg/L for maintenance (in liquid culture media) | ||
-'''[[User:Michael S. Siddiqui|Michael S. Siddiqui]] 21:53, 4 January 2011 (EST)'''* 01/04/2011 (based mainly on communications from [[User:Andrewb|Andrew Babiskin]]) | -'''[[User:Michael S. Siddiqui|Michael S. Siddiqui]] 21:53, 4 January 2011 (EST)'''* 01/04/2011 (based mainly on communications from [[User:Andrewb|Andrew Babiskin]]) | ||
Revision as of 20:42, 17 January 2011
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 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 selection:
- Must be in YP media (doesn't work with YNB)
- 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)
