IGEM:Virginia 2012/Protocols/Bordet Gengou (BG) Agar Plate Preparation
This protocol makes the Bordet Gengou Agar Plates used for optimizing the isolation of Bordatella.
- 30g of Bordet Gengou Agar Base
- 1 L of distilled H2O
- 13 mL glycerol
- 2600 mL Fernback Flask
- Stir Bar
- 100 mL screw cap bottles
- Autoclave tape
- 4°C Chamber
- Add 30g of Bordet Gengou Agar Base to 1 liter of distilled H2O in a 2600 mL Fernback Flask while stirring.
- Turn on heat to medium hot.
- Add 13 mL of glycerol
- When thoroughly heated and in solution, aliquot 80 mL in to 100 mL screw cap bottles.
- Leave caps loose enough for air exchange and place autoclave tape between cap and bottle.
- Tighten the caps and store at 4°C when done.
- Heat autoclaved BG agar bottles in hot water bath until agar is completely liquid.
- This requires hottest setting on hot plate. Remove bottles to cool on bench top.
- When comfortable to touch, add 13 mL defibrinated sheep blood and pour into 60 x 15 mm culture dishes, avoiding air bubbles. (If blood is added when BG agar is too hot, it will hemolyze the blood, turning it brown).
- Note: for antibiotic plates, add 1 mL of 100x solution of antibiotic at time blood is added.
- When plates have cooled and become solid, turn them upside down.
- Leave overnight to reduce condensation, then package, date, label, and store at 4°C.
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