Team:Columbia-Cooper-NYC/Making Solid Media

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(Difference between revisions)
(Solid Media Protocol)
(Solid Media Protocol)
Line 206: Line 206:
<BLOCKQUOTE>
<BLOCKQUOTE>
Autoclave solution of:
Autoclave solution of:
-
<UL>500 mL deionized water
+
<UL>
 +
<LI>500 mL deionized water
<LI>27 g (NH4)2SO4
<LI>27 g (NH4)2SO4
<LI>0.9 g KCl
<LI>0.9 g KCl

Revision as of 03:50, 4 October 2012


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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