Team:Columbia-Cooper-NYC/Making Solid Media

From 2012.igem.org


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

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