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Solid Media Protocol

Solution A

Sterile filter solution with:

• 2 g 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 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
• 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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