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Team:Kyoto/Protocol
From 2011.igem.org
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* NH4Cl : 1 g | * NH4Cl : 1 g | ||
* 100 mM MgSO4 : 10 ml | * 100 mM MgSO4 : 10 ml | ||
| - | * 20% glucose : 10 mL | + | * 20 % glucose : 10 mL |
* 10 mM CaCl2 : 10 ml | * 10 mM CaCl2 : 10 ml | ||
* 100 mM thiamine-HCl : 10 ml | * 100 mM thiamine-HCl : 10 ml | ||
| - | * 20% casamino acid : 10 ml | + | * 20 % casamino acid : 10 ml |
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# Stir Na2HPO4, KH2PO4, NaCl and NH4Cl with water. | # 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. | + | # After autoclave, add 10 ml filter sterilized 100 mM MgSO4, 20 % glucose, 10 mM CaCl2, 100 mM thiamine-HCl. |
| - | # If you need, add 10ml filter sterilized 20% casamino acid. | + | # If you need, add 10ml filter sterilized 20 % casamino acid. |
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<br/> | <br/> | ||
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===SOB medium=== | ===SOB medium=== | ||
::{| class="wikitable" | ::{| class="wikitable" | ||
Revision as of 05:25, 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 : 500mL
- dry yeast : 20g
- corn flour : 45g
- glucose : 50g
- agarose : 3.5~5g
- propionic acid : 1.5mL
- 10% p-hydroxybenzoate in 70% Eternol : 5g
- 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 10ml filter sterilized 20 % casamino acid.
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.
