|
|
(92 intermediate revisions not shown) |
Line 1: |
Line 1: |
- | {{Kyoto_Foreground}} | + | {{Kyoto_Foreground|active_page=method}} |
| {{Kyoto_Background}} | | {{Kyoto_Background}} |
| {{Kyoto_WikiDesign}} | | {{Kyoto_WikiDesign}} |
| | | |
- | <!-- main -->
| + | ='''Protocol'''= |
- | <div id="main"> | + | Listed below our protocol for this project.<br> |
| + | We hope you find them useful!! |
| | | |
- | = '''Protocol''' = | + | <html><a name="cloning"></a></html> |
| | | |
- | === Medium for drosophila ===
| + | [[File:Cloning.PNG|link=https://2011.igem.org/Team:Kyoto/Cloning|Cloning]]<br> |
- | ::{| class="wikitable" | + | [[File:Medium.PNG|link=https://2011.igem.org/Team:Kyoto/Medium|Medium]]<br> |
- | |+ [http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Medium for drosophila]
| + | [[File:Measurement.PNG|link=https://2011.igem.org/Team:Kyoto/Measurement|Measurement]] |
- | ! width="300" |Materials
| + | |
- | ! width="300" |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.■
| + | |
- | |}
| + | |
- | <br/> | + | |
- | ===M9 medium===
| + | |
- | ::{| class="wikitable" | + | |
- | ! width="300" |Materials
| + | |
- | ! width="300" |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.
| + | |
- | |}
| + | |
- | <br/> | + | |
- | ===SOB medium===
| + | |
- | ::{| class="wikitable" | + | |
- | ! width="300" |Materials
| + | |
- | ! width="300" |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.
| + | |
- | |}
| + | |
- | <br/>
| + | |
- | | + | |
- | ===SOC medium===
| + | |
- | ::{| class="wikitable" | + | |
- | ! width="300" |Materials
| + | |
- | ! width="300" |Methods
| + | |
- | |-
| + | |
- | |
| + | |
- | * SOB : 100 ml
| + | |
- | * 2 M glucose: 1 ml
| + | |
- | |
| + | |
- | # Add 2 M glucose to 100 ml SOB.
| + | |
- | |}
| + | |
- | | + | |
- | </div>
| + | |
- | <!-- end of main -->
| + | |
Listed below our protocol for this project.
We hope you find them useful!!