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Template:Kyoto/Notebook/Sep 18
From 2013.igem.org
Contents |
Sep 18
Restriction Enzyme Digestion
| 9/18Plux | SpeI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 3.2µL | 1µL | 1µL | 3µL | 3µL | 18.8µL | 30µL |
| NC | 1.7µL | 0µL | 0µL | 1µL | 1µL | 6.3µL | 10µL |
| 9/18Pcon-pT181antisense-spinach-DT② | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 7.5µL | 1µL | 1µL | 3µL | 3µL | 14.5µL | 30µL |
| NC | 1.4icro;L | 0µL | 0µL | 1µL | 1µL | 6.6µL | 10µL |
| 9/16 Plac② | SpeI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 13.8µL | 1µL | 1µL | 3µL | 3µL | 14.5µL | 30µL |
| NC | 0.7µL | 0µL | 0µL | 1µL | 1µL | 7.3µL | 10µL |
| 9/16 Spinach-DT | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 17.6µL | 1µL | 1µL | 3µL | 3µL | 4.4µL | 30µL |
| NC | 0.9µL | 0µL | 0µL | 1µL | 1µL | 7.1µL | 10µL |
| 9/8 Pcon J23100 | SpeI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 5.7µL | 1µL | 1µL | 3µL | 3µL | 16.3µL | 30µL |
| NC | 0.3µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL |
| 9/16 RBS-lacZα-DT | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 8.4µL | 1µL | 1µL | 3µL | 3µL | 13.6µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
| 9/11 Pcon-pT181antisense-spinach-DT①: | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 7.2µL | 1µL | 1µL | 3µL | 3µL | 14.8µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
| 9/6 apt12-1R-DT | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 7.9µL | 1µL | 1µL | 3µL | 3µL | 14.1µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
| 9/3 pT181antisense | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 6.3µL | 1µL | 1µL | 3µL | 3µL | 15.7µL | 30µL |
| NC | 0.3µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL |
| 9/14 pSB4K5 | EcoRI | SpeI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 7.3µL | 1µL | 1µL | 3µL | 3µL | 14.7µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
| 8/31 Plux-RBS-GFP-DT | EcoRI | SpeI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 9.2µL | 1µL | 1µL | 3µL | 3µL | 12.8µL | 30µL |
| NC | 0.5µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL |
| 9/12 RBS-lysis3-DT | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 5.2µL | 1µL | 1µL | 3µL | 3µL | 16.8µL | 30µL |
| NC | 0.3µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
| 9/18 Pcon-pT181attenuator | EcoRI | SpeI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2cuts | 14.3µL | 1µL | 1µL | 3µL | 3µL | 7.7µL | 30µL |
| NC | 0.7µL | 0µL | 0µL | 1µL | 1µL | 7.3µL | 10µL |
Electrophoresis
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | Plae② | Spel | Ptsl |
| 2 | RBS-lysis3-DT | Xbal | Ptsl |
| 3 | Plux-RBS-GFP-DT | EcoR1 | Spel |
| 4 | PSB4K5 | EcoR1 | Spel |
| 5 | Pcon | Spel | Pstl |
| 6 | Plux | Spel | Pstl |
| 7 | 1kbp ladder | -- | -- |
| 8 | Plae② | -- | -- |
| 9 | RBS-lysis3-DT | -- | -- |
| 10 | Plux-RBS-GFP-DT | -- | -- |
| 11 | PSB4K5 | -- | -- |
| 12 | Pcon | -- | -- |
| 13 | Plux | -- | -- |
Electrophoresis
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | 100bp ladder | -- | -- |
| 2 | Pcon pT181antisense spinach-DT② | Xbal | Pstl |
| 3 | Spinach-DT | Xbal | Pstl |
| 4 | RBS-lacZα-DT | Xbal | Pstl |
| 5 | Pcon pT181antisense-Spinach-DT① | Xbal | Pstl |
| 6 | apt12-1R-DT | Xbal | Pstl |
| 7 | pT181attenuator | Xbal | Pstl |
| 8 | Pcon attenuator | EcoR1 | Spel |
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | 100bp ladder | -- | -- |
| 2 | Pcon pT181antisense spinach-DT② | Xbal | Pstl |
| 3 | Spinach-DT | Xbal | Pstl |
| 4 | RBS-lacZα-DT | Xbal | Pstl |
| 5 | Pcon pT181antisense-Spinach-DT① | Xbal | Pstl |
| 6 | apt12-1R-DT | Xbal | Pstl |
| 7 | pT181attenuator | Xbal | Pstl |
| 8 | Pcon attenuator | EcoR1 | Spel |
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | 100bp ladder | -- | -- |
| 2 | Pcon pT181antisense spinach-DT② | -- | -- |
| 3 | Spinach-DT | -- | -- |
| 4 | RBS-lacZα-DT | -- | -- |
| 5 | Pcon pT181antisense-Spinach-DT① | -- | -- |
| 6 | apt12-1R-DT | -- | -- |
| 7 | pT181attenuator | -- | -- |
| 8 | Pcon attenuator | -- | -- |
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kbp ldder | -- |
| 2 | Plac② | SpeI+PstI |
| 3 | Plac② | SpeI+PstI |
| 4 | -- | -- |
| 5 | RBS-lysis3-DT | XbaI+PstI |
| 6 | RBS-lysis3-DT | XbaI+PstI |
| 7 | -- | -- |
| 8 | Plux-RBS-GFP-DT | EcoRI+SpeI |
| 9 | Plux-RBS-GFP-DT | EcoRi+SpeI |
| 10 | -- | -- |
| 11 | PSB4KS | EcoRI+SpeI |
| 12 | PSB4KS | EcoRI+SpeI |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kbp ldder | -- |
| 2 | Pcon | SpeI+PstI |
| 3 | Pcon | SpeI+PstI |
| 4 | -- | -- |
| 5 | Plux | SpeI+PStI |
| 6 | Plux | SpeI+PStI |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | Pcon PT181 antisense spinach-DT | XbaI+PstI |
| 2 | Pcon PT181 antisense Spinach-DT | XbaI+PstI |
| 3 | 100bp ladder | -- |
| 4 | Pcon PT181 antisense Spinach-DT | XbaI+PstI |
| 5 | Pcon PT181 antisense Spinach-DT | XbaI+PstI |
| 6 | 1kbp ladder | -- |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 2 | apt12-1R-DT | XbaI+PstI |
| 3 | apt12-1R-DT | XbaI+PstI |
| 4 | -- | -- |
| 5 | PT181 attenuator | XbaI+PstI |
| 6 | PT181 attenuator | XbaI+PstI |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 2 | Pcoa attenuator | EcoRI+PstI |
| 3 | Pcoa attenuator | EcoRI+PstI |
| 4 | -- | -- |
| 5 | RBS-lacZα-DT | XbaI+PstI |
| 6 | RBS-lacZα-DT | XbaI+PstI |
