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Team:Hawaii/Protocols/Solid growth media
From 2008.igem.org
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==BG-11== | ==BG-11== | ||
===Materials=== | ===Materials=== | ||
| - | + | ||
* 940 ml nanopure H<sub>2</sub>O | * 940 ml nanopure H<sub>2</sub>O | ||
* 20 ml 50X thiosulfate (final conc. of 1mM) | * 20 ml 50X thiosulfate (final conc. of 1mM) | ||
* 20 ml 50X TES buffer, pH 8.0 (final conc. of 10mM) | * 20 ml 50X TES buffer, pH 8.0 (final conc. of 10mM) | ||
| + | * 20 ml 50X BG-11 | ||
* 15 g agar | * 15 g agar | ||
Revision as of 23:22, 24 June 2008
Contents |
LB agar
Materials
- 25 g LB broth mixture (Difco)
- 15 g agar
- 1 L ddH2O
Procedure
- Mix dry ingredients together with 1 L ddH2O using a magnetic stir bar.
- Autoclave to sterilize. Let cool to 55C.
- Pour 28 ml LB solution for thin plates; 50 ml for thick plates.
LB + amp100
Materials
- 1 L LB agar
- 1 ml 1000X ampicillin stock solution
Procedure
- Make LB agar as directed.
- When mixture is cooled to 55C, add 1 ml 1000X ampicilllin stock solution.
- Plate as usual.
LB+kan20+amp50
Materials
- 1 L LB agar
- 0.5 mL 1000X ampicillin stock solution
- 0.4 mL 1000X kanamycin stock solution
Procedure
- Make LB as directed.
- After cooling to 55C, add ampicillin and kanamycin.
- Plate as usual.
LB+sp100
Materials
- 1 L LB agar
- 1.0 mL 1000X spectinomycin stock solution
Procedure
- Make LB as directed.
- Cool to 55C, then add spectinomycin.
- Plate as usual.
BG-11
Materials
- 940 ml nanopure H2O
- 20 ml 50X thiosulfate (final conc. of 1mM)
- 20 ml 50X TES buffer, pH 8.0 (final conc. of 10mM)
- 20 ml 50X BG-11
- 15 g agar
Procedure
- Combine agar and nanopure H2O. Stir mixture with a magnetic stir bar.
- Autoclave to sterilize. Cool to 55C.
- Add TES, thiosulfate, and BG-11
- Test pH (should be ~8.0)
- Plate 28 ml for thin plates; 50 mL for thick plates.
