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Team:Kyoto/Protocol
From 2011.igem.org
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# After autoclave, add 10 ml filter sterilized 100 mM MgSO4, 20 % glucose, 10 mM CaCl2, 100 mM thiamine-HCl. | # After autoclave, add 10 ml filter sterilized 100 mM MgSO4, 20 % glucose, 10 mM CaCl2, 100 mM thiamine-HCl. | ||
# If you need, add 10 ml filter sterilized 20 % casamino acid. | # If you need, add 10 ml filter sterilized 20 % casamino acid. | ||
| + | |} | ||
| + | <br/> | ||
| + | |||
| + | ===LB medium=== | ||
| + | ::{| class="wikitable" | ||
| + | ! width="300" |Materials | ||
| + | ! width="300" |Methods | ||
| + | |- | ||
| + | | | ||
| + | * water : 1 L | ||
| + | * Tryptone: 10 g | ||
| + | * Yeast extract : 5 g | ||
| + | * NaCl : 10 g | ||
| + | * Agar : 15g | ||
| + | | | ||
| + | # Stir Tryptone, Yeast extract and NaCl with water. | ||
| + | # If you make LB plates, add agar. | ||
| + | # Autoclave. | ||
|} | |} | ||
<br/> | <br/> | ||
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# Stir Tryptone and Yeast extract with water. | # Stir Tryptone and Yeast extract with water. | ||
| - | # Add 200 ul 5 M NaCl and 84 ul 3 M KCl | + | # Add 200 ul 5 M NaCl and 84 ul 3 M KCl. |
# After autoclave, add 1 ml filter sterilized 10 mM MgSO4 and 10 mM MgCl2. | # After autoclave, add 1 ml filter sterilized 10 mM MgSO4 and 10 mM MgCl2. | ||
|} | |} | ||
Revision as of 05:30, 23 September 2011
Contents |
Protocol
Medium for drosophila
[http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Medium for drosophila] Materials Methods - water : 500 mL
- dry yeast : 20 g
- corn flour : 45 g
- glucose : 50 g
- agarose : 3.5~5 g
- propionic acid : 1.5 mL
- 10 % p-hydroxybenzoate in 70 % Eternol : 5 g
- Stir dry yeast and agarose with about two-thirds of water. Then, autoclave it.
- Stir corn flour and glucose with the remaining water.
- Stir 1 and 2, then autoclave it again.
- after autoclave, add propionic acid and 10 % p-hydroxybenzoate in 70 % Eternol into it.■
M9 medium
Materials Methods - water : 1 L
- Na2HPO4: 6 g
- KH2PO4 : 3 g
- NaCl : 0.5 g
- NH4Cl : 1 g
- 100 mM MgSO4 : 10 ml
- 20 % glucose : 10 mL
- 10 mM CaCl2 : 10 ml
- 100 mM thiamine-HCl : 10 ml
- 20 % casamino acid : 10 ml
- Stir Na2HPO4, KH2PO4, NaCl and NH4Cl with water.
- After autoclave, add 10 ml filter sterilized 100 mM MgSO4, 20 % glucose, 10 mM CaCl2, 100 mM thiamine-HCl.
- If you need, add 10 ml filter sterilized 20 % casamino acid.
LB medium
Materials Methods - water : 1 L
- Tryptone: 10 g
- Yeast extract : 5 g
- NaCl : 10 g
- Agar : 15g
- Stir Tryptone, Yeast extract and NaCl with water.
- If you make LB plates, add agar.
- Autoclave.
SOB medium
Materials Methods - water : 100 mL
- Tryptone: 2 g
- Yeast extract : 0.5 g
- 5 M NaCl : 200 ul
- 3 M KCl : 84 ul
- 1 M MgSO4 : 1 ml
- 1 M MgCl2 : 1 ml
- Stir Tryptone and Yeast extract with water.
- Add 200 ul 5 M NaCl and 84 ul 3 M KCl.
- After autoclave, add 1 ml filter sterilized 10 mM MgSO4 and 10 mM MgCl2.
SOC medium
Materials Methods - SOB : 100 ml
- 2 M glucose: 1 ml
- Add 2 M glucose to 100 ml SOB.
