Smolke Endy LB Agar

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(Recipe)
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Day before, ask how much is needed to make.
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Day before, ask how much is needed to make. Usually 8 500ml bottles.
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Log the batch number in your notebook.
Log the batch number in your notebook.
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From recipe, measure Bacto-peptone, Yeast, NaCl, and Agar into each bottle.  
From recipe, measure Bacto-peptone, Yeast, NaCl, and Agar into each bottle.  
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Add 450ml of water from the millipore filter in the washroom and add a stir bar.
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Add 500ml of water from the millipore filter in the lab using the automatic measure.  
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Mix on the stirplate.
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Remove stirbar with stirbar retriever and pour most of the mixture into a graduated cylinder. As you get to the bottom of the bottle, swirl the mixture and pour. Bring up to final volume of 500ml and pour back into the original bottle. Swirl and pour as you transfer to make sure as much of the mixture is returned to the bottle.
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Adhere autoclave tape. Loosely cap bottle and set in autoclave tray with 2 inches of water.  
Adhere autoclave tape. Loosely cap bottle and set in autoclave tray with 2 inches of water.  
Just prior to closing the door, give each bottle another swirl to ensure mixture has not settled out.
Just prior to closing the door, give each bottle another swirl to ensure mixture has not settled out.
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Autoclave on Liquid 60.  
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Autoclave on Liquid 500.  
Individually remove each bottle and gently swirl contents. Let cool, secure cap closed and store in the weigh room cabinet.
Individually remove each bottle and gently swirl contents. Let cool, secure cap closed and store in the weigh room cabinet.
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*2.55 G NaCl
*2.55 G NaCl
*6.1 G Agar
*6.1 G Agar
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* Add 500ml of Milli-Q water or DI water and sterilize for 45 min.
 

Revision as of 13:46, 16 May 2012

Day before, ask how much is needed to make. Usually 8 500ml bottles.

Log the batch number in your notebook.

Label each bottle with LB Agar, lot number, and your initials.

Make each 500ml in a 1 Liter bottle. Since Agar will not go into solution at room temp. It would be difficult to pour the mixture into aliquots from a larger batch.

From recipe, measure Bacto-peptone, Yeast, NaCl, and Agar into each bottle. Add 500ml of water from the millipore filter in the lab using the automatic measure.


Adhere autoclave tape. Loosely cap bottle and set in autoclave tray with 2 inches of water. Just prior to closing the door, give each bottle another swirl to ensure mixture has not settled out.

Autoclave on Liquid 500.

Individually remove each bottle and gently swirl contents. Let cool, secure cap closed and store in the weigh room cabinet.

Recipe

per 500ml bottle

  • 5.1 G Bacto Peptone
  • 2.55 G Yeast Extract
  • 2.55 G NaCl
  • 6.1 G Agar
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