IGEM:Virginia 2012/Protocols/Bordet Gengou (BG) Agar Plate Preparation: Difference between revisions
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(New page: ==Materials== * 30g of Bordet Gengou Agar Base * 1 L of distilled H<sub>2</sub>O * 13 mL glycerol * 2600 mL Fernback Flask * Stir Bar * 100 mL screw cap bottles * Autoclave * Autoclave ta...) |
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* Leave caps loose enough for air exchange and place autoclave tape between cap and bottle. | * Leave caps loose enough for air exchange and place autoclave tape between cap and bottle. | ||
* Autoclave | * Autoclave | ||
* Tighten the caps and store at | * Tighten the caps and store at 4°C when done. | ||
===Pour plates=== | ===Pour plates=== | ||
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* ''Note: for antibiotic plates, add 1 mL of 100x solution of antibiotic at time blood is added.'' | * ''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'''. | * When plates have cooled and become solid, '''turn them upside down'''. | ||
* Leave overnight to reduce condensation, then package, date, label, and store at | * Leave overnight to reduce condensation, then package, date, label, and store at 4°C. | ||
==Notes== | ==Notes== | ||
Revision as of 10:57, 31 May 2012
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 4°C 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 4°C.
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
References
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.
