Team:Columbia-Cooper-NYC/Making Solid Media

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== Solid Media Protocol ==
== Solid Media Protocol ==
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'''Solution A'''
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<BLOCKQUOTE>
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Sterile filter solution with:
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<UL>
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<LI>2 g of Na2S2O3
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<LI>10 mL of H2O
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</UL>
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</BLOCKQUOTE>
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'''Solution B'''
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<BLOCKQUOTE>
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Autoclave solution of (2.5% tryptone):
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<UL>
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<LI>2.5g tryptone broth
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<LI>100 mL of H2O
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</UL>
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</BLOCKQUOTE>
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'''Solution C'''
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<BLOCKQUOTE>
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Sterile filter solution of:
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<UL>
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<LI>500 mL deionized water
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<LI>27 g (NH4)2SO4
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<LI>0.9 g KCl
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<LI>1.605 g leucine
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<LI>4.5 g MgSO4
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<LI>0.02 g HK2PO4
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<LI>0.485 g diaminopimelic acid
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<LI> ~0.7 mL H2SO4 (or enough to bring pH down to ~2.0)
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<LI>20 g Fe(II)SO4 (after adding everything else above)
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</UL>
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</BLOCKQUOTE>
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'''Solution D'''
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<BLOCKQUOTE>
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Autoclave solution containing (4% agarose):
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<UL>
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<LI>20 g Agarose
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<LI>500 mL distilled water
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</UL>
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</BLOCKQUOTE>
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<OL>
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<LI>Heat up agarose solution until it becomes liquid
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<LI>Pick a volume of solid media to make
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<LI>Mix 1% vol A & 50% vol D, and 1% vol B & 48% vol C
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<LI>Mix together AD and BC
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<LI>Pour into petri dishes and allow to harden
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</OL>
<i>Credit for protocol goes to Jason Candreva, Research Associate, Columbia University</i>
<i>Credit for protocol goes to Jason Candreva, Research Associate, Columbia University</i>
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Return to [[Team:Columbia-Cooper-NYC/Protocols|Protocols Page]]

Latest revision as of 03:01, 27 October 2012


Solid Media Protocol

Solution A

Sterile filter solution with:
  • 2 g of Na2S2O3
  • 10 mL of H2O

Solution B

Autoclave solution of (2.5% tryptone):
  • 2.5g tryptone broth
  • 100 mL of H2O

Solution C

Sterile filter solution of:
  • 500 mL deionized water
  • 27 g (NH4)2SO4
  • 0.9 g KCl
  • 1.605 g leucine
  • 4.5 g MgSO4
  • 0.02 g HK2PO4
  • 0.485 g diaminopimelic acid
  • ~0.7 mL H2SO4 (or enough to bring pH down to ~2.0)
  • 20 g Fe(II)SO4 (after adding everything else above)

Solution D

Autoclave solution containing (4% agarose):
  • 20 g Agarose
  • 500 mL distilled water
  1. Heat up agarose solution until it becomes liquid
  2. Pick a volume of solid media to make
  3. Mix 1% vol A & 50% vol D, and 1% vol B & 48% vol C
  4. Mix together AD and BC
  5. Pour into petri dishes and allow to harden

Credit for protocol goes to Jason Candreva, Research Associate, Columbia University

Return to Protocols Page

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