MicroStation System, MicroLog Version 4.2 - DTU Systems Biology ...

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Technical Notes
General hints for culture media preparation
Keep dehydrated media powder in original bottles with lids tightly closed
to avoid water absorption and deterioration.
Use clean dry glassware that has been rinsed free of all soap residue.
Add water to the vessel first, then weigh agar powder and add it to the
vessel. Mix to obtain an even suspension. Do NOT fill the vessel more
than two-thirds full (to avoid boiling over during heating and autoclaving).
Heat agar gently, with constant stirring.
Making inoculating fluid (GN/GP-IF)
Biolog supplies pre-made inoculating fluid, with the following
composition:
• 0.40% sodium chloride (NaCl)
• 0.03% Pluronic F-68 (e.g., Sigma P7061)
• 0.02% Gellan Gum (e.g., Phytagel, Sigma P8169)
However, if you wish to prepare your own, using Biolog’s formula.
1. Add 0.2 grams Gellan Gum to 1 liter of H2O.
2. Heat to boiling, stirring constantly, until the Gellan Gum is completely
dissolved.
3. Turn off the heat but continue stirring.
4. Add 4 grams NaCl and continue stirring until the NaCl is completely
dissolved.
5. Add 0.3 grams of Pluronic F-68. Continue stirring until the Pluronic F-
68 is completely dissolved.
6. Dispense the volume appropriate to obtain 19 ml post-autoclaving into
20 x 150 mm tubes.
7. Sterilize by autoclaving at 15 pounds of pressure and 121° C for 30
minutes.
Note: Gellan Gum concentration of 0.01-0.02% is acceptable.
MicroStation System/MicroLog User Guide Apr 07 Section 9 � Page 5