Team:Caltech/Recipes

From 2011.igem.org

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{{Team:Caltech/templateheader|
{{Team:Caltech/templateheader|
Content=
Content=
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'''50% glycerol Stock:'''
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* 50 mL glycerol
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* 50 mL nanopure water
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<br/>
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'''Agar/LB plate (Autoclaved):'''  
'''Agar/LB plate (Autoclaved):'''  
* 500 mL nanopure water
* 500 mL nanopure water
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<br/>  
<br/>  
-
'''50% glycerol Stock:'''  
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'''Ampicillin Stock'''
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* 50 mL glycerol
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*Mass out amount needed for 50 mg/ml or 100 mg/ml stock of ampicillin trihydrate
-
* 50 mL nanopure water  
+
*Add a small amount of nanopure water
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*Add 1M NaOH until solution turns clear (we are making ampicillin sodium salt in this step, which is much more soluble in water)
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*Fill to total volume with nanopure water}}
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<br/>  
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'''Chloramphenicol Stock'''
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*for 25 mg/ml stock solution (usually, chloramphenicol is used at 12.5 µg/ml or 25 µg/ml)
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*add 250 mg chloramphenicol to 10 ml 100% ethanol
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*store at -20°C
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<br/>
<p>'''Enrichment Minimal Media'''</p>   
<p>'''Enrichment Minimal Media'''</p>   
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* variable amounts of EDC's
* variable amounts of EDC's
<br/>
<br/>
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'''Gibson Mix(1.33x):'''
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* 50 μl of Taq Ligase
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* 100 μl of 5x isothermal buffer
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* 2 μl of T5 exconuclease
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* 5 μl of Phusion polymerase
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* Nuclease-free water
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* 15 μl of Master Mix?
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<br/>
<p>'''SOC Media'''</p>
<p>'''SOC Media'''</p>
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*after cooling below 50 degrees Celsius, add 5 mL filter-sterilized 20% glucose solution.
*after cooling below 50 degrees Celsius, add 5 mL filter-sterilized 20% glucose solution.
<br/>
<br/>
-
 
-
'''Chloramphenicol Stock'''
 
-
*for 25 mg/ml stock solution (usually, chloramphenicol is used at 12.5 µg/ml or 25 µg/ml)
 
-
*add 250 mg chloramphenicol to 10 ml 100% ethanol
 
-
*store at -20°C
 
-
<br/>
 
-
 
-
'''Ampicillin Stock'''
 
-
*Mass out amount needed for 50 mg/ml or 100 mg/ml stock of ampicillin trihydrate
 
-
*Add a small amount of nanopure water
 
-
*Add 1M NaOH until solution turns clear (we are making ampicillin sodium salt in this step, which is much more soluble in water)
 
-
*Fill to total volume with nanopure water}}
 

Revision as of 22:47, 14 July 2011


Caltech iGEM 2011



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50% glycerol Stock:
  • 50 mL glycerol
  • 50 mL nanopure water


Agar/LB plate (Autoclaved):

  • 500 mL nanopure water
  • 10 g tryptone
  • 10 g NaCl
  • 5 g yeast extract
  • 15 g agar


Ampicillin Stock

  • Mass out amount needed for 50 mg/ml or 100 mg/ml stock of ampicillin trihydrate
  • Add a small amount of nanopure water
  • Add 1M NaOH until solution turns clear (we are making ampicillin sodium salt in this step, which is much more soluble in water)
  • Fill to total volume with nanopure water



Chloramphenicol Stock

  • for 25 mg/ml stock solution (usually, chloramphenicol is used at 12.5 µg/ml or 25 µg/ml)
  • add 250 mg chloramphenicol to 10 ml 100% ethanol
  • store at -20°C


Enrichment Minimal Media

  • 1.0712 g K2 HPO4
  • 0.5239 g KH2 PO4
  • 79 mL Phosphate solution
  • 10 mL Salt Solution I
  • 10 mL Salt Solution II
  • 1 mL Wolfe's Vitamin Solution
  • 0.1 mL SL-10 Trace Element Solution.
  • variable amounts of EDC's


Gibson Mix(1.33x):

  • 50 μl of Taq Ligase
  • 100 μl of 5x isothermal buffer
  • 2 μl of T5 exconuclease
  • 5 μl of Phusion polymerase
  • Nuclease-free water
  • 15 μl of Master Mix?


SOC Media

  • for 250 ml (adapted from [http://openwetware.org/wiki/SOC OpenWetWare])
  • 1.25 g yeast extract
  • 5 g tryptone
  • 0.146 g NaCl
  • 0.0465 g KCl
  • 0.616 g MgSO4•7H2O
  • 0.508 g MgCl2•6H2O
  • adjust pH to 7.5 using 1M NaOH
  • autoclave
  • after cooling below 50 degrees Celsius, add 5 mL filter-sterilized 20% glucose solution.


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