Team:Alberta/Notebook/protocols/LB

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==General Protocols==
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/invitro_biobyte_assembly | In Vitro BioByte Assembly]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/LB | LB Plates and Broth]]
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*[[Team:Alberta/Notebook/protocols/transformations |Transformations]]
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*[[Team:Alberta/Notebook/protocols/overnight |5mL Overnight ]]
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*[[Team:Alberta/Notebook/protocols/glycerol | Glycerol Stock ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/miniprep | Plasmid Miniprep ]]
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*[[Team:Alberta/Notebook/protocols/digest | Restriction Digest ]]
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*[[Team:Alberta/Notebook/protocols/vector_dephos | Vector Dephosphorylation]]
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*[[Team:Alberta/Notebook/protocols/ligation | Ligation ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/agarose_gel | Agarose Gel Electrophoresis ]]
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*[[Team:Alberta/Notebook/protocols/gel_extraction | Gel Extraction ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/pcr | PCR]]
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*[[Team:Alberta/Notebook/protocols/colony_pcr | Colony PCR ]]
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*[[Team:Alberta/Notebook/protocols/pcr_purification | PCR Purification ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/labelling | Sample Labelling Conventions]]
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*[[Team:Alberta/Notebook/protocols/sequencing | Fluorescent Sequencing Reaction]]
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*[[Team:Alberta/Notebook/protocols/primer_design | Primer Design]]
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==Protocol: Making LB Plates==
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==Making LB Plates==
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===Procedure:===
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===LB Agar (To Make 1L):===
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LB Agar (To Make 1L):
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*Before you start:
*Before you start:
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** DON`T put tubes of antibiotics back into freezer.
** DON`T put tubes of antibiotics back into freezer.
* Add the following to 800 ml MilliQ H20:
* Add the following to 800 ml MilliQ H20:
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{|
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::Bacto-tryptone || 10g
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|Bacto-tryptone || 10g
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::Yeast extract || 5g
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|-
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::NaCl || 10g
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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).
* Adjust pH to 7.5 with NaOH (Optional).
* Add 15g agar.
* Add 15g agar.
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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.  
* 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.
* Pour plates, making sure they are color coded with the type of antibiotic that you used.
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==Making LB Broth:==
==Making LB Broth:==
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Latest revision as of 02:21, 27 October 2010

TEAM ALBERTA


Making LB Plates

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.