Team:Columbia-Cooper-NYC/Making Solid Media

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

Retrieved from "http://2012.igem.org/Team:Columbia-Cooper-NYC/Making_Solid_Media"