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==Yeast Nitrogen Base Minus Amino Acids (YNB-AA) Recipe==
 
==Yeast Nitrogen Base Minus Amino Acids (YNB-AA) Recipe==
 
*For 1 L Batch:
 
*For 1 L Batch:
 
**1.7g Yeast Nitrogen Base (stored in cold room)
 
**1.7g Yeast Nitrogen Base (stored in cold room)
 
**5g NH3SO4 (Ammonium Sulfate, stored in chemical stock cabinet)
 
**5g NH3SO4 (Ammonium Sulfate, stored in chemical stock cabinet)
 
+
*Dissolve in 800mL Np H2O for YNB-AA stock
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)
 
Autoclave (store at RT for media stocks)
  
*To use as working media:
+
'''To use as working media:'''
 
*After autoclaving add:
 
*After autoclaving add:
 
**100mL 10X (20%) Dextrose  
 
**100mL 10X (20%) Dextrose  
**100mL 10X Drop Out stock (10X DO stock recipe: [[http://openwetware.org/wiki/Smolke:Protocols/DO_Media]])
+
**100mL 10X Drop Out stock (10X DO stock recipe here: [[http://openwetware.org/wiki/Smolke:Protocols/DO_Media]])
 
+
**1 L Total Volume
**1 L Total
 
  
 
==Yeast Nitrogen Base Minus Amino Acids Non-Inducing, Non-Repressing (YNB-AA NINR) Recipe==
 
==Yeast Nitrogen Base Minus Amino Acids Non-Inducing, Non-Repressing (YNB-AA NINR) Recipe==
For 1 L Batch:
+
*For 1 L Batch:
1.7g Yeast Nitrogen Base
+
**1.7g Yeast Nitrogen Base
5g NH3SO4  
+
**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)
  
Dissolve in 700mL Np H2O for YNB-AA NINR stock
+
'''To use as working media:'''
Autoclave (store at RT for media stocks)
+
*After autoclaving add:
 
+
**100mL 10X (20%) Raffinose
To use as working media:
+
**100mL 10X (10%) Sucrose
After autoclaving add:
+
**100mL 10X Drop Out stock
100mL 10X (20%) Raffinose
+
**1 L Total Volume
100mL 10X (10%) Sucrose
 
 
 
100mL 10X Drop Out stock
 
 
 
1 L Total
 
  
 
==Yeast Peptone (YP) Media Recipe==
 
==Yeast Peptone (YP) Media Recipe==
For 1 L Batch:
+
*For 1 L Batch:
10g Yeast Extract [Bacto™ REF#212750] (stored in chemical stock cabinet)
+
**10g Yeast Extract [Bacto™ REF#212750] (stored in chemical stock cabinet)
20g Peptone [Bacto™ REF#211677] (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
+
**Dissolve in 900mL Np H2O for YP stock
 
Autoclave (store at RT for media stocks)
 
Autoclave (store at RT for media stocks)
 
+
'''To use as working media:'''
To use as working media:
 
 
After autoclaving add:
 
After autoclaving add:
100mL 10X (20%) Dextrose
+
**100mL 10X (20%) Dextrose
1 L Total YPD (Yeast Peptone Dextrose)
+
**1 L Total Volume YPD (Yeast Peptone Dextrose)
  
To pour any of these media stocks as plates:
+
'''To pour any of these media stocks as plates:'''
Add 20g agar per 1L final media volume before autoclaving
+
**Add 20g agar per 1L final media volume before autoclaving
See here [[http://openwetware.org/wiki/Smolke:Protocols/Dropout_plates]]
+
**See here [[http://openwetware.org/wiki/Smolke:Protocols/Dropout_plates]]
  
For yeast KanMX selection:
+
'''For yeast KanMX (G418) selection:'''
Add 400mg/L for selection (in plates)
+
*Must be in YP media (doesn't work with YNB)
Add 200mg/L for maintenance (in liquid culture media)
+
*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)
  
-'''[[User:Michael S. Siddiqui|Michael S. Siddiqui]] 21:53, 4 January 2011 (EST)'''* 01/04/2011 (based mainly on communications from 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 00:32, 14 October 2011

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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)