From 2008.igem.org
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| - | == List of Minipreps === | + | == Stock Glycerol === |
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| | + | === Protocol === |
| | + | * 1mL glycerol |
| | + | * 1mL LB |
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| | == Transformations == | | == Transformations == |
Revision as of 10:33, 5 August 2008
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← Yesterday ↓ Calendar ↑Tomorrow →
souche stockage
MiniPreps
Conditions
- Use of Quiagen's protocol on all the clones cultivated on the 31th.
- Preparation of 50µl of minipreps in water.
Stock Glycerol =
Protocol
| Name
| Biobrick
| Description
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| MP L102
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| MP L103
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| MP L104
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| MP L105
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| MP L106
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| MP L107
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| MP L108
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| MP L110
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| MP L111
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| MP L112
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| MP L115
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| MP L116
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| MP L117
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| MP L118
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| MP L119
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| MP L121
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| MP L122
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| MP L124
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| MP L125
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Transformations
Protocol
use of TOP10 Chemically competent cells
- Defroze competent cells on ice during 5'
- Add 5µl of DNA Ligation in 50µL of competent bacterias (or 1µL for the positive control puc19)
- Incubate 30' on ice
- Heat-shock the cells during 30" at 42°C without shaking
- Put 2' on ice
- Add 250µL of pre-warmed SOC medium (4°C)
- Incubate 1h at 37°C under shaking (225rpm)
- Spin at 5.000rpm during 30"
- Remove 150µL of supernatant
- Resuspent the pellet in the 150µL left
- Spread on adequated plates
- Incubate O/N at 37°C
List of transformation
| Name
| Biobrick
| Description
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| L100
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| RBS-TetR-ECFP
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| L101
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| RBS-tetR-GFP tripart
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| L113
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| pBAD-ECFP
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| L114
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| pBAD-GFP
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| L120
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| tetR-ECFP
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| L122
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| RBS-lasI-ECFP
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| L123
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| RBS-lasI-GFP-ter
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| L126
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| RBS-lasR-LVA
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"
