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Team:Alberta/Notebook/protocols/LB
From 2010.igem.org
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** DON`T put tubes of antibiotics back into freezer. | ** DON`T put tubes of antibiotics back into freezer. | ||
* Add the following to 800 ml MilliQ H20: | * Add the following to 800 ml MilliQ H20: | ||
| - | + | ::Bacto-tryptone || 10g | |
| - | + | ::Yeast extract || 5g | |
| - | + | ::NaCl || 10g | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
* Adjust pH to 7.5 with NaOH (Optional). | * Adjust pH to 7.5 with NaOH (Optional). | ||
* Add 15g agar. | * Add 15g agar. | ||
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* Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder. | * Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder. | ||
* Pour plates, making sure they are color coded with the type of antibiotic that you used. | * Pour plates, making sure they are color coded with the type of antibiotic that you used. | ||
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==Making LB Broth:== | ==Making LB Broth:== | ||
Revision as of 17:57, 26 October 2010
Protocol: Making LB Plates
Procedure:
LB Agar (To Make 1L):
- Before you start:
- Use AGAR not agarose!
- Use an Erlenmeyer flask that is twice the volume of your solution (avoid spillage in autoclave).
- Keep your flask on a stir plate with a stir rod.
- Put Aluminum foil on the top of the flask and put autoclave tape on.
- DON`T let it solidify after autoclaving by keeping it on a stir plate while it cools.
- DON`T put tubes of antibiotics back into freezer.
- Add the following to 800 ml MilliQ H20:
- Bacto-tryptone || 10g
- Yeast extract || 5g
- NaCl || 10g
- Adjust pH to 7.5 with NaOH (Optional).
- Add 15g agar.
- Melt agar into solution in the microwave (Optional).
- Adjust volume to 1L with MilliQ H20
- Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
- Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.
- Pour plates, making sure they are color coded with the type of antibiotic that you used.
Making LB Broth:
- All of the above WITHOUT AGAR.
