Team:WashU/Protocols/LBPlates

From 2012.igem.org

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<!--Thanks to UCLA and Kansas State University for the above instructions.  The original source can be found here: http://www.eeb.ucla.edu/Faculty/Barber/Web%20Protocols/LB%20Agar%20Plates.pdf http://www.k-state.edu/hermanlab/protocols/AntibioticUsage.htm-->
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<!--Thanks to UCLA and Kansas State University for the above instructions.  The original source can be found here: http://www.eeb.ucla.edu/Faculty/Barber/Web%20Protocols/LB%20Agar%20Plates.pdf http://www.k-state.edu/hermanlab/protocols/AntibioticUsage.htm
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{{WashUback}}
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NOTE:We have modified the protocol slightly to suit our purposes.-->
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{{WashUbackprotocols}}
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<div id = "protocolshort">
=LB plate=
=LB plate=
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#Autoclave mixture for 20 minutes on liquid cycle.
#Autoclave mixture for 20 minutes on liquid cycle.
#Allow mixture to cool to around 55 degrees Celsius.
#Allow mixture to cool to around 55 degrees Celsius.
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#(optional). add appropriate amount of antibiotic. Final concentrations are given. Ampicillin-100ug/ml, Kanamycin-50ug/ml, Tetracycline-15ug/ml, Chloramphenicol-50ug/ml
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#Pour 15 ml of agar mixture into plates, and allow to solidify. (The number of mls poured depends on how thick you want your plates to be - to have thicker plates, pour more into each plate.)
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#Use, or store in refrigerator.
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(optional). add appropriate amount of antibiotic. Final concentrations are given. Ampicillin-100ug/ml, Kanamycin-50ug/ml, Tetracycline-15ug/ml, Chloramphenicol-100ug/ml
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NOTE: to make liquid lb media. Follow all the above directions except do not add agar.
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<div align="center">
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#Pour 25 ml of agar mixture into plates, and allow to solidify.
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<font size ="5">
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#Use, or store in refrigerator.
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[https://2012.igem.org/Team:WashU/Protocols Back to Protocols]

Latest revision as of 21:32, 9 August 2012


LB plate

Making the plate

  1. Mix 25 grams of LB broth powder and 10 grams of agar with 1 Liter of deionized water.
  2. Shake the container until all of the agar and broth powder dissolves.
  3. Autoclave mixture for 20 minutes on liquid cycle.
  4. Allow mixture to cool to around 55 degrees Celsius.
  5. (optional). add appropriate amount of antibiotic. Final concentrations are given. Ampicillin-100ug/ml, Kanamycin-50ug/ml, Tetracycline-15ug/ml, Chloramphenicol-50ug/ml
  6. Pour 15 ml of agar mixture into plates, and allow to solidify. (The number of mls poured depends on how thick you want your plates to be - to have thicker plates, pour more into each plate.)
  7. Use, or store in refrigerator.

NOTE: to make liquid lb media. Follow all the above directions except do not add agar.

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