Protocol/15

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Protocol 15: Making LB Plates
Protocol 15: Making LB Plates
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Makes 1L (~50 plates)
 
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Notes:  
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Procedure:
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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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1.Add the following to 800 ml MilliQ H20
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*10g Bacto-tryptone
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*5g yeast extract
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*10g NaCl
+
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2.Adjust pH to 7.5 with NaOH - optional
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LB Agar (To Make 1L):
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3.Add 15g agar  
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*Before you start:
 +
** Use AGAR not agarose!
 +
** Use an Erlenmeyer flask that is twice the volume of your solution (avoid spillage in autoclave).
 +
** Keep your flask on a stir plate with a stir rod.
 +
** Put Aluminum foil on the top of the flask and put autoclave tape on.
 +
** DON`T let it solidify after autoclaving by keeping it on a stir plate while it cools.
 +
** DON`T put tubes of antibiotics back into freezer.
 +
* Add the following to 800 ml MilliQ H20:
 +
{|
 +
|Bacto-tryptone || 10g
 +
|-
 +
|Yeast extract || 5g
 +
|-
 +
|NaCl || 10g
 +
|}
 +
* Adjust pH to 7.5 with NaOH (Optional).
 +
* Add 15g agar.
 +
* Melt agar into solution in the microwave (Optional).
 +
* Adjust volume to 1L with MilliQ H20
 +
* Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
 +
* Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.
 +
* Pour plates, making sure they are color coded with the type of antibiotic that you used.
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4.Melt agar into solution in the microwave - optional
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Making LB Broth:
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5.Adjust volume to 1L with MilliQ H20
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*All of the above WITHOUT AGAR.
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6.Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
 
-
 
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7.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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8.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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Revision as of 01:49, 26 October 2010

TEAM ALBERTA

Protocol 15: Making LB Plates


Procedure:

LB Agar (To Make 1L):

  • Before you start:
    • Use AGAR not agarose!
    • Use an Erlenmeyer flask that is twice the volume of your solution (avoid spillage in autoclave).
    • Keep your flask on a stir plate with a stir rod.
    • Put Aluminum foil on the top of the flask and put autoclave tape on.
    • DON`T let it solidify after autoclaving by keeping it on a stir plate while it cools.
    • DON`T put tubes of antibiotics back into freezer.
  • Add the following to 800 ml MilliQ H20:
Bacto-tryptone 10g
Yeast extract 5g
NaCl 10g
  • Adjust pH to 7.5 with NaOH (Optional).
  • Add 15g agar.
  • Melt agar into solution in the microwave (Optional).
  • Adjust volume to 1L with MilliQ H20
  • Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
  • Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.
  • Pour plates, making sure they are color coded with the type of antibiotic that you used.

Making LB Broth:

  • All of the above WITHOUT AGAR.


Back

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