Team:Columbia-Cooper-NYC/Making Solid Media

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(Solid Media Protocol)
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== Solid Media Protocol ==
== Solid Media Protocol ==
 +
'''Solution A'''
 +
<BLOCKQUOTE>
 +
Sterile filter solution with:
 +
<UL>
 +
<LI>2g of Na2S2O3
 +
<LI>10 mL of H2O
 +
</UL>
 +
</BLOCKQUOTE>
 +
'''Solution B'''
 +
<BLOCKQUOTE>
 +
Adjust pH to 2.0 with sulfuric acid (H2SO4) and sterile filter solution with:
 +
<UL>
 +
<LI>Fe(II)SO4
 +
<LI>10 mL of H2O
 +
</UL>
 +
</BLOCKQUOTE>
 +
'''Solution C'''
 +
<BLOCKQUOTE>
 +
Autoclave solution of:
 +
<UL>500 mL deionized
 +
<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>10 g Fe(II)SO4 (at the end)
 +
</UL>
 +
</BLOCKQUOTE>
 +
 +
'''Solution D'''
 +
<BLOCKQUOTE>
 +
Autoclave solution containing:
 +
<UL>
 +
<LI>6 g Agarose
 +
<LI>480 mL distilled water
 +
</UL>
 +
</BLOCKQUOTE>
 +
 +
<OL>
 +
<LI>Mix A & D, and B & 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]]

Revision as of 03:47, 4 October 2012


Solid Media Protocol

Solution A

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

Solution B

Adjust pH to 2.0 with sulfuric acid (H2SO4) and sterile filter solution with:
  • Fe(II)SO4
  • 10 mL of H2O

Solution C

Autoclave solution of:
    500 mL deionized
  • 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
  • 10 g Fe(II)SO4 (at the end)

Solution D

Autoclave solution containing:
  • 6 g Agarose
  • 480 mL distilled water
  1. Mix A & D, and B & C
  2. Mix together AD and BC
  3. Pour into petri dishes and allow to harden

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

Return to Protocols Page