Smolke:Protocols/Yeast Media Recipes: Difference between revisions

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==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)
 
**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 selection:'''
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 Andrew Babiskin)
-'''[[User:Michael S. Siddiqui|Michael S. Siddiqui]] 21:53, 4 January 2011 (EST)'''* 01/04/2011 (based mainly on communications from Andrew Babiskin)

Revision as of 20:01, 4 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:

    • 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)