Team:Columbia-Cooper-NYC/Making Solid Media

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(→Solid Media Protocol)
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Adjust pH to 2.0 with sulfuric acid (H2SO4) and sterile filter solution with:
Adjust pH to 2.0 with sulfuric acid (H2SO4) and sterile filter solution with:
<UL>
<UL>
-
<LI>Fe(II)SO4
+
<LI>2 g Fe(II)SO4
<LI>10 mL of H2O
<LI>10 mL of H2O
</UL>
</UL>

Revision as of 03:49, 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:
  • 2 g 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

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