Lethbridge/Notebook
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+ | ==July 07 2007== | ||
+ | Attempted Transformation of 4 Biobricks | ||
+ | 1. BBa_J5526 - Plate 3 well 6F - RFP | ||
+ | 2. BBa_I13522 - Plate 2 well 15H - GFP | ||
+ | 3. BBa_R0011 - Plate 1 well 7M - LacI inhibited promoter | ||
+ | 4. BBa_PO440 - Plate 1 well 21K - GFP repressor | ||
+ | |||
+ | Protocol | ||
+ | 1. Add 15uL deionized H2O to well | ||
+ | 2. Add 1 uL plasmid to 25uL DH5alpha | ||
+ | 3. Incubate on ice for 30min | ||
+ | 4. Heat shock without shaking for 20sec | ||
+ | 5. Place on ice for 2min | ||
+ | 6. Add 0.5mL LB, incubate for 1hr | ||
+ | 7. Plate 100uL on amp+ LB plates, incubate overnight at 37 C | ||
+ | |||
+ | |||
+ | |||
===July 24=== | ===July 24=== | ||
Note book created | Note book created |
Revision as of 17:14, 25 August 2007
Contents |
July 07 2007
Attempted Transformation of 4 Biobricks
1. BBa_J5526 - Plate 3 well 6F - RFP 2. BBa_I13522 - Plate 2 well 15H - GFP 3. BBa_R0011 - Plate 1 well 7M - LacI inhibited promoter 4. BBa_PO440 - Plate 1 well 21K - GFP repressor
Protocol
1. Add 15uL deionized H2O to well 2. Add 1 uL plasmid to 25uL DH5alpha 3. Incubate on ice for 30min 4. Heat shock without shaking for 20sec 5. Place on ice for 2min 6. Add 0.5mL LB, incubate for 1hr 7. Plate 100uL on amp+ LB plates, incubate overnight at 37 C
July 24
Note book created
July 25
* GFP repressor (TetR) and LacI inhibited promoter transformed successfully * RFP with promoter, RBS, and terminators transformed but does not emit RF under UV light and plasmid too small, will have to build from five subparts. * GFP, LacI, and four subparts of RFP left to go
July 27
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