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Protocol/15
From 2010.igem.org
Protocol 15: Making LB Plates
Makes 1L (~50 plates)
Notes:
- 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.
1.Add the following to 800 ml MilliQ H20
- 10g Bacto-tryptone
- 5g yeast extract
- 10g NaCl
2.Adjust pH to 7.5 with NaOH - optional
3.Add 15g agar
4.Melt agar into solution in the microwave - optional
5.Adjust volume to 1L with MilliQ H20
6.Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
7.Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.
8.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.
