From 2013.igem.org
Sep 18
Restriction Enzyme Digestion
No name
| | 9/18Plux | Spel | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 18.8µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 6.3µL | 10µL
|
| 9/18Pcon-pT181antisense-spin-DT② | Xbal | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 14.5µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 6.6µL | 10µL
|
| | 9/16 Plac② | Spel | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 14.5µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.3µL | 10µL
|
| | 9/16 spin-DT | Xbal | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 4.4µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.1µL | 10µL
|
| | 9/8 Pcon J23100 | Spel | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 16.3µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 9/16 RBS-lacZα-DT | Xbal | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 13.6µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | 9/11 Pcon-pT181antisense-spin-DT①: | Xbal | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 14.8µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | 9/6 apt12-1R-DT | Xbal | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 14.1µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | 9/3 pT181antisense | Xbal | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 15.7µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 9/14 pSB4K5 | EcoR1 | Spel | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 14.7µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | 8/31 Plux-RBS-GFP-DT | EcoR1 | Spel | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 12.8µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL
|
| | 9/12 RBS-lysis3-DT | Xbal | Ptsl | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 16.8µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | 9/18 Pcon-pT181attenuator | EcoR1 | Spel | Buffer | BSA | MilliQ | total
|
| 2cuts | 1µL | 1µL | 3µL | 3µL | 7.7µL | 30µL
|
| NC | 0µL | 0µL | 1µL | 1µL | 7.3µL | 10µL
|
Electrophoresis
No name
| 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 | -- | --
|
File:Igku xxxxxx.xxx
Electrophoresis
Okazaki
| 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 | -- | --
|
File:Igku xxxxxx.xxx
No name
| Lane | DNA | Enzyme
|
| 1 | 1kbp ldder | --
|
| 2 | Plac② | Spel+Pstl
|
| 3 | Plac② | Spel+Pstl
|
| 4 | -- | --
|
| 5 | RBS-lysis3-DT | Xbal+Pstl
|
| 6 | RBS-lysis3-DT | Xbal+Pstl
|
| 7 | -- | --
|
| 8 | Plux-RBS-GFP-DT | EcoR1+Spel
|
| 9 | Plux-RBS-GFP-DT | EcoR1+Spel
|
| 10 | -- | --
|
| PSB4KS | EcoR1+Spel
|
| PSB4KS | EcoR1+Spel
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Lane | DNA | Enzyme
|
| 1 | 1kbp ldder | --
|
| 2 | Pcon | Spel+Pstl
|
| 3 | Pcon | Spel+Pstl
|
| 4 | -- | --
|
| 5 | Plux | Spel+PStl
|
| 6 | Plux | Spel+PStl
|
| Lane | DNA | Enzyme
|
| 1 | Pcon PT181 antisense spinach-DT | Xbal+Pstl
|
| 2 | Pcon PT181 antisense Spinach-DT | Xbal+Pstl
|
| 3 | 100bp ladder | --
|
| 4 | Pcon PT181 antisense Spinach-DT | Xbal+Pstl
|
| 5 | Pcon PT181 antisense Spinach-DT | Xbal+Pstl
|
| 6 | 1kbp ladder | --
|
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder | --
|
| 2 | apt12-1R-DT | Xbal+Pstl
|
| 3 | apt12-1R-DT | Xbal+Pstl
|
| 4 | -- | --
|
| 5 | PT181 attenuator | Xbal+Pstl
|
| 6 | PT181 attenuator | Xbal+Pstl
|
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder | --
|
| 2 | Pcoa attenuator | EcoR1+Pstl
|
| 3 | Pcoa attenuator | EcoR1+Pstl
|
| 4 | -- | --
|
| 5 | RBS-lacZα-DT | Xbal+Pstl
|
| 6 | RBS-lacZα-DT | Xbal+Pstl
|
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