IGEM:Virginia 2012/Protocols/Bordet Gengou (BG) Agar Plate Preparation

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Contents

Materials

  • 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
  • Autoclave tape
  • 4oC Chamber

Procedure

Making solution

  • 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.
  • Autoclave
  • Tighten the caps and store at 4oC when done.

Pour plates

  • 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 4oC.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!


References

Contact

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