Protocol/15

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Protocol 15: Making LB Plates
 
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Procedure:
 
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LB Agar (To Make 1L):
 
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*Before you start:
 
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** Use AGAR not agarose!
 
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** Use an Erlenmeyer flask that is twice the volume of your solution (avoid spillage in autoclave).
 
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** Keep your flask on a stir plate with a stir rod.
 
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** Put Aluminum foil on the top of the flask and put autoclave tape on.
 
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** DON`T let it solidify after autoclaving by keeping it on a stir plate while it cools.
 
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** DON`T put tubes of antibiotics back into freezer.
 
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* Add the following to 800 ml MilliQ H20:
 
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{|
 
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|Bacto-tryptone || 10g
 
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|-
 
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|Yeast extract || 5g
 
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|-
 
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|NaCl || 10g
 
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|}
 
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* Adjust pH to 7.5 with NaOH (Optional).
 
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* Add 15g agar.
 
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* Melt agar into solution in the microwave (Optional).
 
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* Adjust volume to 1L with MilliQ H20
 
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* Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
 
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* Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.
 
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* Pour plates, making sure they are color coded with the type of antibiotic that you used.
 
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Making LB Broth:
 
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*All of the above WITHOUT AGAR.
 
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Latest revision as of 17:13, 26 October 2010