Team:Brown-Stanford/Lab/Protocols/BG11

From 2011.igem.org

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<html><h1>Protocol Code: M1</h1></html>
== '''BG-11''' ==
== '''BG-11''' ==
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''(Taken from [http://microbiology.ucdavis.edu/meeks/BG11medium.html|UC Davis])''
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''(Taken from <html><a href=""http://microbiology.ucdavis.edu/meeks/BG11medium.html"">Meeks Lab at University of California, Davis</a></html>)''  
=== '''Stock solutions for BG-11''' ===
=== '''Stock solutions for BG-11''' ===
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Stock 1:
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==== '''Stock 1''' ====
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0.1g/liter Na2MG EDTA
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*0.1g/liter Na<sub>2</sub>MG EDTA
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0.6g/liter Ferric ammonium citrate
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*0.6g/liter Ferric ammonium citrate
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Citric acid&#8226;1H2O
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*0.6g/liter Citric acid&#8226;1H<sub>2</sub>O
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0.6g/liter
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*3.6g/liter CaCl<sub>2</sub>&#8226;2H<sub>2</sub>O
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CaCl2 . 2H2O
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Filter sterilize into a sterile bottle or autoclave.
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3.6g/liter
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Filter sterilize into a sterile bottle or autoclave
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Stock 2:
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MgSO4 . 7H2O
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7.5g/liter
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Filter sterilize into a sterile bottle or autoclave
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Stock 3:
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K2HPO4 . 3H2O
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4.0g/liter
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or K2HPO4
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3.05g/liter
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Filter sterilize into a sterile bottle or autoclave
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Stock 5 (Microelements):
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H3BO3
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2.86g/liter
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MnCl2 . 4H2O
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1.81g/liter
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ZnSO4 . 7H2O
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0.222g/liter
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CuSO4 . 5H2O
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0.079g/liter
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COCl2 . 6H2O
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0.050g/liter
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NaMoO4 . 2H2O
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0.391g/liter
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or MoO4 (85%)
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0.018g/liter
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==== '''Stock 2''' ====
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*7.5g/liter MgSO<sub>4</sub>&#8226;7H<sub>2</sub>O
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Filter sterilize into a sterile bottle or autoclave.
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==== '''Stock 3''' ====
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*4.0g/liter K<sub>2</sub>HPO<sub>4</sub>&#8226;3H<sub>2</sub>O or 3.05g/liter K<sub>2</sub>HPO<sub>4</sub>
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Filter sterilize into a sterile bottle or autoclave.
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==== '''Stock 5 (Microelements)''' ====
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Stock 5 (Microelements):
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*2.86g/liter H<sub>3</sub>BO<sub>3</sub>
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*1.81g/liter MnCl<sub>2</sub>&#8226;4H<sub>2</sub>O
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*0.222g/liter ZnSO<sub>4</sub>&#8226;7H<sub>2</sub>O
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*0.079g/liter CuSO<sub>4</sub>&#8226;5H<sub>2</sub>O
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*0.050g/liter COCl<sub>2</sub>&#8226;6H<sub>2</sub>O
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*0.391g/liter NaMoO<sub>4</sub>&#8226;2H<sub>2</sub>O or 0.018g/liter MoO<sub>4</sub> (85%)
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B) For basic BG11 medium combine the following stock solutions:
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=== '''Basic BG11 medium''' ===
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Stock Solution Per Liter of medium
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Combine the following stock solutions (for 1 liter):
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Stock 1
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*10 mL Stock 1
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10 ml
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*10 mL Stock 2
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Stock 2
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*10 mL Stock 3
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10 ml
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*0.02 g Na<sub>2</sub>CO<sub>3</sub>
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Stock 3
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*1.0 mL Stock 5
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10 ml
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*1.5 g NaNO<sub>3</sub>
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Na2CO3
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0.02g
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Stock 5
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1.0 ml
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NaNO3
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1.5g
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Combine stocks and adjust pH to 7.5 (use 1.0N HCl). Aliquot into flasks (50 ml/125 ml flask) with cotton stoppers on top and autoclave. After autoclaving and cooling the pH is about 7.1. For solid media add 1% noble agar. For BG-11o don't add NaNO3.
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Combine stocks and adjust pH to 7.5 (use 1.0N HCl). Aliquot into flasks (50 mL/125 mL flask) with cotton stoppers on top and autoclave. After autoclaving and cooling the pH is about 7.1. For solid media add 1% noble agar. For BG-11, don't add NaNO<sub>3</sub>.
{{:Team:Brown-Stanford/Templates/Foot}}
{{:Team:Brown-Stanford/Templates/Foot}}

Latest revision as of 18:15, 28 September 2011

Brown-Stanford
iGEM

Protocol Code: M1

BG-11

(Taken from Meeks Lab at University of California, Davis)

Stock solutions for BG-11

Stock 1

  • 0.1g/liter Na2MG EDTA
  • 0.6g/liter Ferric ammonium citrate
  • 0.6g/liter Citric acid•1H2O
  • 3.6g/liter CaCl2•2H2O

Filter sterilize into a sterile bottle or autoclave.

Stock 2

  • 7.5g/liter MgSO4•7H2O

Filter sterilize into a sterile bottle or autoclave.

Stock 3

  • 4.0g/liter K2HPO4•3H2O or 3.05g/liter K2HPO4

Filter sterilize into a sterile bottle or autoclave.

Stock 5 (Microelements)

Stock 5 (Microelements):

  • 2.86g/liter H3BO3
  • 1.81g/liter MnCl2•4H2O
  • 0.222g/liter ZnSO4•7H2O
  • 0.079g/liter CuSO4•5H2O
  • 0.050g/liter COCl2•6H2O
  • 0.391g/liter NaMoO4•2H2O or 0.018g/liter MoO4 (85%)

Basic BG11 medium

Combine the following stock solutions (for 1 liter):

  • 10 mL Stock 1
  • 10 mL Stock 2
  • 10 mL Stock 3
  • 0.02 g Na2CO3
  • 1.0 mL Stock 5
  • 1.5 g NaNO3

Combine stocks and adjust pH to 7.5 (use 1.0N HCl). Aliquot into flasks (50 mL/125 mL flask) with cotton stoppers on top and autoclave. After autoclaving and cooling the pH is about 7.1. For solid media add 1% noble agar. For BG-11, don't add NaNO3.