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Team:Hawaii/Protocols/Solid growth media
From 2008.igem.org
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(→BG-11 + 5% LB) |
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===Materials=== | ===Materials=== | ||
* 25 g LB broth mixture (Difco) | * 25 g LB broth mixture (Difco) | ||
| + | * 15 g agar | ||
* 1 L ddH<sub>2</sub>O | * 1 L ddH<sub>2</sub>O | ||
| Line 11: | Line 12: | ||
== LB + amp<sub>100</sub>== | == LB + amp<sub>100</sub>== | ||
===Materials=== | ===Materials=== | ||
| - | 1 L LB agar | + | * 1 L LB agar |
| - | 1 ml 1000X [[Team:Hawaii/Protocols/Antibiotics|ampicillin stock solution]] | + | * 1 ml 1000X [[Team:Hawaii/Protocols/Antibiotics|ampicillin stock solution]] |
| - | + | ===Procedure=== | |
# Make LB agar as directed. | # Make LB agar as directed. | ||
# When mixture is cooled to 55C, add 1 ml 1000X ampicilllin stock solution. | # When mixture is cooled to 55C, add 1 ml 1000X ampicilllin stock solution. | ||
| Line 20: | Line 21: | ||
==LB+kan<sub>20</sub>+amp<sub>50</sub>== | ==LB+kan<sub>20</sub>+amp<sub>50</sub>== | ||
===Materials=== | ===Materials=== | ||
| - | 1 L LB agar | + | * 1 L LB agar |
| - | 0.5 mL 1000X [[Team:Hawaii/Protocols/Antibiotics|ampicillin stock solution]] | + | * 0.5 mL 1000X [[Team:Hawaii/Protocols/Antibiotics|ampicillin stock solution]] |
| + | * 0.4 mL 1000X [[Team:Hawaii/Protocols/Antibiotics|kanamycin stock solution]] | ||
| + | |||
| + | ===Procedure=== | ||
| + | # Make LB as directed. | ||
| + | # After cooling to 55C, add ampicillin and kanamycin. | ||
| + | # Plate as usual. | ||
| + | |||
| + | ==LB+sp<sub>100</sub>== | ||
| + | ===Materials=== | ||
| + | * 1 L LB agar | ||
| + | * 1.0 mL 1000X [[Team:Hawaii/Protocols/Antibiotics|spectinomycin stock solution]] | ||
| + | |||
| + | === Procedure=== | ||
| + | # Make LB as directed. | ||
| + | # Cool to 55C, then add spectinomycin. | ||
| + | # Plate as usual. | ||
| + | |||
| + | ==BG-11 agar== | ||
| + | ===Materials=== | ||
| + | |||
| + | * 940 ml nanopure H<sub>2</sub>O | ||
| + | * 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 H<sub>2</sub>O. 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. | ||
| + | |||
| + | ==BG-11 + 5% LB== | ||
| + | ===Materials=== | ||
| + | * 1.25 g LB broth mixture (Difco) | ||
| + | * 15 g agar | ||
| + | * 940 ml nanopure H<sub>2</sub>O | ||
| + | * 20 ml 50X BG-11 | ||
| + | * 20 ml 50X thiosulfate | ||
| + | * 20 ml 50X TES buffer | ||
| + | ===Procedure=== | ||
| + | # Combine LB and agar with the nanopure H<sub>2</sub>O. | ||
| + | # Autoclave to sterilize. | ||
| + | # Add TES, thiosulfate, and BG-11. | ||
| + | # Test pH (~8.0). | ||
| + | # Plate as usual. | ||
| + | |||
| + | ==BG-11+sp<sub>2.5</sub>+sm<sub>2.5</sub>== | ||
| + | ===Materials=== | ||
| + | * BG-11 agar | ||
| + | * 0.15 ml 1000X [[Team:Hawaii/Protocols/Antibiotics|spectinomycin stock solution]] | ||
| + | * 0.3 ml 1000X [[Team:Hawaii/Protocols/Antibiotics|streptomycin stock solution]] | ||
| + | |||
| + | ===Procedure=== | ||
| + | # Make BG-11 agar as directed. | ||
| + | # After adding TES, thiosulfate, and BG-11, add spectinomycin and streptomycin. | ||
| + | # Plate as usual. | ||
Latest revision as of 01:50, 25 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 agar
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.
BG-11 + 5% LB
Materials
- 1.25 g LB broth mixture (Difco)
- 15 g agar
- 940 ml nanopure H2O
- 20 ml 50X BG-11
- 20 ml 50X thiosulfate
- 20 ml 50X TES buffer
Procedure
- Combine LB and agar with the nanopure H2O.
- Autoclave to sterilize.
- Add TES, thiosulfate, and BG-11.
- Test pH (~8.0).
- Plate as usual.
BG-11+sp2.5+sm2.5
Materials
- BG-11 agar
- 0.15 ml 1000X spectinomycin stock solution
- 0.3 ml 1000X streptomycin stock solution
Procedure
- Make BG-11 agar as directed.
- After adding TES, thiosulfate, and BG-11, add spectinomycin and streptomycin.
- Plate as usual.
