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

Making Biolog Universal Growth Agar (BUG)
1. Mix the following in a 2-3 liter flask:
• 57 grams BUG Agar
• 1,000 ml purified, distilled, or deionized water
Technical Notes
2. Gently boil while stirring to dissolve the agar and other components.
3. Cool an aliquot and measure the pH. Adjust pH with NaOH or HCl to
achieve a final pH of 7.3 + 0.1.
4. Sterilize by autoclaving at 15 pounds of pressure, at 121° C, for 15
minutes.
5. Cool to 45-50° C.
6. Dispense into sterile petri dishes.
Making Biolog Universal Anaerobe Agar + blood (BUA + B)
1. Mix the following in a 2-3 liter container:
• 51.7 grams BUA Agar
• 950 ml purified, distilled, or deionized water
2. While flushing with oxygen-free nitrogen gas, gently boil mixture
while stirring to dissolve the agar and other components.
3. Cool an aliquot and measure the pH. Adjust pH with NaOH or HCl to
achieve a final pH of 7.2 + 0.1 at 25° C.
4. Sterilize by autoclaving at 15 pounds of pressure and 121° C for 15
minutes. Make sure the bottle is tightly capped to prevent the entry of
oxygen.
5. Cool to 45-50° C under a stream of oxygen-free nitrogen gas.
6. Add 50 ml sterile fresh defibrinated sheep’s blood just prior to
dispensing and mix gently. (Use good quality blood with a hematocrit
of at least 40%).
7. Dispense into sterile petri dishes in an anaerobic chamber.
MicroStation System/MicroLog User Guide Apr 07 Section 9 � Page 3