From 2013.igem.org
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Revision as of 07:53, 24 September 2013
Sep 5
Colony PCR not yet
No name
| Sample | base pair
|
| 8/9 J23100-3 | 1142
|
| 8/9 J23100-4 | 1142
|
| 8/9 J23100-5 | 1142
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5m | 30s | 30s | 30s | 30cycles
|
Electrophoresis
No name
| Lane | Sample | Enzyme1 | Enzyme2
|
| 1 | 100bp ladder | -- | --
|
| 2 | J23100-3 | -- | --
|
| 3 | J23100-4 | -- | --
|
| 4 | J23100-5 | -- | --
|
| 5 | 100bp ladder | -- | --
|
File:Igku xx.xxx
Miniprep
No name
| DNA | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-pT181 anti | 243.0 | 1.90 | 1.81
|
| Pcon-Spinach-DT | 253.7 | 1.57 | 1.51
|
Ligation
Kojima
| state | Vector | Inserter | Ligation High ver.2
|
| experiment | Pconst | 13.2 | spinach-DT | 2.1 | 3.5
|
| experiment | Plac | 15.3 | spinach-DT | 2.1 | 3.5
|
| experiment | Pconst | 13.2 | pT181 antisense (XbaI & PstI) | 6.6 | 3.5
|
| experiment | Plac | 15.3 | pT181 antisense (XbaI & PstI) | 6.6 | 3.5
|
| experiment | Ptet | 30.1 | pT181 antisense (XbaI & PstI) | 6.6 | 3.5
|
| experiment | Pconst | 13.2 | pT181 attenuator | 11.7 | 3.5
|
| experiment | Plac | 15.3 | pT181 attenuator | 11.7 | 3.5
|
| experiment | Ptet | 30.1 | RBS-lacZα-DT | 3.0 | 3.5
|
incubate 16 °C 1 hour
No name
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder |
|
| 2 | RBS-lysis1-DT | XbaI & PstI
|
| 3 | RBS-lysis1-DT | XbaI & PstI
|
| 5 | RBS-lysis3-DT | XbaI & PstI
|
| 6 | RBS-lysis3-DT | XbaI & PstI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| RBS-lysis1-DT (XbaI & PstI) | 5.3 | 1.98 | 0.06
|
| RBS-lysis3-DT (XbaI & PstI) | 5.3 | 1.98 | 0.06
|
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder |
|
| 3 | Pcon-pT181 attenuator | EcoRI & SpeI
|
| 4 | Pcon-pT181 attenuator | EcoRI & SpeI
|
| 6 | Spinach-DT | XbaI & PstI
|
| 7 | Spinach-DT | XbaI & PstI
|
| 9 | RBS-lacZα-DT | XbaI & PstI
|
| 10 | RBS-lacZα-DT | XbaI & PstI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-pT181 attenuator (EcoRI & SpeI) | 5.2 | 1.82 | 0.36
|
| Spinach-DT (XbaI & PstI) | 8.0 | 1.87 | 0.25
|
| RBS-lacZα-DT (XbaI & PstI) | 10.3 | 1.71 | 0.54
|
| Lane | DNA | Enzyme
|
| 1 | 1kbp ladder |
|
| 2 | Ptet-pT181 antisense | EcoRI & PstI
|
| 3 | Ptet-pT181 antisense | EcoRI & PstI
|
| 5 | pSB4K5 | EcoRI & SpeI
|
| 6 | pSB4K5 | EcoRI & SpeI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Ptet-pT181 antisense(EcoRI & PstI) | 30.6 | 1.84 | 1.16
|
| pSB4K5 (EcoRI & SpeI) | 21.8 | 1.87 | 0.98
|
| Lane | DNA | Enzyme
|
| 1 | 1kbp ladder |
|
| 2 | Pcon-pT181 attenuator | SpeI & PstI
|
| 3 | Pcon-pT181 attenuator | SpeI & PstI
|
| 5 | Plac | SpeI & PstI
|
| 6 | Plac | SpeI & PstI
|
| 8 | pSB1C3 | EcoRI & SpeI
|
| 9 | pSB1C3 | EcoRI & SpeI
|
| 11 | pSB1C3 | XbaI & PstI
|
| 12 | pSB1C3 | XbaI & PstI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-pT181 attenuator (SpeI & PstI) | 14.5 | 1.91 | 1.09
|
| Plac (SpeI & PstI) | 11.6 | 1.74 | 1.19
|
| pSB1C3 (EcoRI & SpeI) | 5.7 | 1.74 | 0.34
|
| pSB1C3 (XbaI & PstI) | 5.6 | 2.00 | 0.37
|
Colony PCR
Hirano
| Sample | base pair
|
| 8/9 BBa_J23100 3 | --
|
| 8/9 BBa_J23100 4 | --
|
| 8/9 BBa_J23100 5 | --
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 30s | 30cycles
|
Liquid Culture
Nakamoto
| Sample | medium
|
| 9/11Fusion6 antisense-1 | Plusgrow medium(+CP)
|
| Plac(BBa-R0011)-3 | Plusgrow medium(+Amp)
|
| Plac(BBa-R0011)-4 | Plusgrow medium(+Amp)
|
| Fusion3m2 attenuator -1 | Plusgrow medium(+CP)
|
| Pλ-luxI-2 | Plusgrow medium(+Amp)
|
| 9/11 aptamer12-1M-2 | Plusgrow medium(+CP)
|
Transformation
じっけんしゃ
| Name | Sample(µL) | Competent Cells(µL) | Total(µL) | Plate
|
| 9/8 Pcont+aptamer 12-1R-DT | 1 | 10 | 11 | Amp
|
| 9/8 Pcon+RBS-lacZα-DT | 1 | 10 | 11 | Amp
|
| 9/8 Pcon-pT181 attenuater+aptamer 12-1R-DT | 1 | 10 | 11 | Amp
|
| 9/8 Plac+RBS-lysis1-DT | 2 | 20 | 22 | CP
|
| 9/8 Plac+RBS-lysis2-DT | 2 | 20 | 22 | CP
|
| 9/8 Plac+aptamer 12-1R-DT | 2 | 20 | 22 | CP
|
| 9/8 Plac+spinach | 2 | 20 | 22 | CP
|
| 9/8 Plac+pT181 attenuater | 2 | 20 | 22 | CP
|
| 9/8 Plux+RBS-lysis1-DT | 1 | 10 | 11 | CP
|
| 9/8 Plux+RBS-lysis2-DT | 1 | 10 | 11 | CP
|
| 9/8 Ptet+RBS-lacZα-DT | 1 | 10 | 11 | CP
|
| 9/8 Ptet-pT181 anti+spinach-DT | 1 | 10 | 11 | CP
|
| 9/8 Pcon-pT181 attenuater+DT | 1 | 10 | 11 | CP
|
Restriction Enzyme Digestion
じっけんしゃ
| | 9/8 Pcon-pT181 anti | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 8.2µL | 0µL | 1µL | 0µL | 1µL | 3µL | 3µL | 13.8µL | 30µL
|
| NC | 0.4µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | 9/8 Pcon-spin-DT | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 7.9µL | 1µL | 1µL | 0µL | 0µL | 3µL | 3µL | 14.1µL | 30µL
|
| NC | 0.4µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | 8/21 F1 atte | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 6.5µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 15.5µL | 30µL
|
| NC | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 8/21 F3m2 atte | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 6.0µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 16µL | 30µL
|
| NC | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 8/21 F1 anti | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 6.7µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 15.3µL | 30µL
|
| NC | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 8/21 F6 anti | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 6.0µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 16µL | 30µL
|
| NC | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 8/21 tetR aptamer 12_1M | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 5.3µL | 1µL | 1µL | 0µL | 0µL | 3µL | 3µL | 16.7µL | 30µL
|
| NC | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 8/21 tetR aptamer 12_P | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 13.3µL | 1µL | 1µL | 0µL | 0µL | 3µL | 3µL | 8.7µL | 30µL
|
| NC | 0.7µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.3µL | 10µL
|
| | 9/4 spin-DT | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 5.4µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 16.6µL | 30µL
|
| NC | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL
|
| | 9/7 RBS-lysis1-DT | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 10µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 12µL | 30µL
|
| NC | 0.5µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL
|
| | 9/6 RBS-lysis2-DT | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 13µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 9µL | 30µL
|
| NC | 0.7µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.3µL | 10µL
|
| | 9/7 RBS-lysis3-DT | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 10µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 12µL | 30µL
|
| NC | 0.5µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL
|
| | 8/20 J23100 RBS-lacZα-DT-(1) | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 7µL | 0.5µL | 0.5µL | 0µL | 0µL | 3µL | 3µL | 16µL | 30µL
|
| NC | 0.5µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL
|
| | 8/20 RBS-lacZα-DT | E | S | X | P | BSA | Buffer | MilliQ | total
|
| 2 cuts | 10µL | 0.5µL | 0.5µL | 0µL | 0µL | 3µL | 3µL | 13µL | 30µL
|
| NC | 0.5µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL
|
5*M9(EDTA)
No name
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder |
|
| 2 | RBS-lysis1-DT | XbaI & PstI
|
| 3 | RBS-lysis1-DT | XbaI & PstI
|
| 5 | RBS-lysis3-DT | XbaI & PstI
|
| 6 | RBS-lysis3-DT | XbaI & PstI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| RBS-lysis1-DT (XbaI & PstI) | 5.3 | 1.98 | 0.06
|
| RBS-lysis3-DT (XbaI & PstI) | 5.3 | 1.98 | 0.06
|
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder |
|
| 3 | Pcon-pT181 attenuator | EcoRI & SpeI
|
| 4 | Pcon-pT181 attenuator | EcoRI & SpeI
|
| 6 | Spinach-DT | XbaI & PstI
|
| 7 | Spinach-DT | XbaI & PstI
|
| 9 | RBS-lacZα-DT | XbaI & PstI
|
| 10 | RBS-lacZα-DT | XbaI & PstI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-pT181 attenuator (EcoRI & SpeI) | 5.2 | 1.82 | 0.36
|
| Spinach-DT (XbaI & PstI) | 8.0 | 1.87 | 0.25
|
| RBS-lacZα-DT (XbaI & PstI) | 10.3 | 1.71 | 0.54
|
| Lane | DNA | Enzyme
|
| 1 | 1kbp ladder |
|
| 2 | Ptet-pT181 antisense | EcoRI & PstI
|
| 3 | Ptet-pT181 antisense | EcoRI & PstI
|
| 5 | pSB4K5 | EcoRI & SpeI
|
| 6 | pSB4K5 | EcoRI & SpeI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Ptet-pT181 antisense(EcoRI & PstI) | 30.6 | 1.84 | 1.16
|
| pSB4K5 (EcoRI & SpeI) | 21.8 | 1.87 | 0.98
|
| Lane | DNA | Enzyme
|
| 1 | 1kbp ladder |
|
| 2 | Pcon-pT181 attenuator | SpeI & PstI
|
| 3 | Pcon-pT181 attenuator | SpeI & PstI
|
| 5 | Plac | SpeI & PstI
|
| 6 | Plac | SpeI & PstI
|
| 8 | pSB1C3 | EcoRI & SpeI
|
| 9 | pSB1C3 | EcoRI & SpeI
|
| 11 | pSB1C3 | XbaI & PstI
|
| 12 | pSB1C3 | XbaI & PstI
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-pT181 attenuator (SpeI & PstI) | 14.5 | 1.91 | 1.09
|
| Plac (SpeI & PstI) | 11.6 | 1.74 | 1.19
|
| pSB1C3 (EcoRI & SpeI) | 5.7 | 1.74 | 0.34
|
| pSB1C3 (XbaI & PstI) | 5.6 | 2.00 | 0.37
|
Miniprep
No name
| DNA | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-pT181 anti | 243.0 | 1.90 | 1.81
|
| Pcon-Spinach-DT | 253.7 | 1.57 | 1.51
|
Colony PCR
No name
| Sample | base pair
|
| 9/8 Plac-spinach-DT-(1) | 659
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5m | 30s | 30s | 40s | 30cycles
|
| Sample | base pair
|
| 9/8 Plac-spinach-DT-(2) | 659
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5m | 30s | 30s | 40s | 30cycles
|
| Sample | base pair
|
| 9/8 Pcon-pt181atte-DT-(1) | 584
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5m | 30s | 30s | 40s | 30cycles
|
Electrophoresis
No name
| Lane | Sample | Enzyme1 | Enzyme2
|
| 1 | 100bp ladder | -- | --
|
| 2 | 8/19 lysis1 | XbaI | PstI
|
| 3 | 8/19 lysis1 | XbaI | --
|
| 4 | 8/19 lysis1 | -- | PstI
|
| 5 | 8/19 lysis1 | -- | --
|
| 6 | 8/19 lysis1(Gel Extraction Product) | XbaI | PstI
|
Liquid Culture
Hirano
| Sample | medium
|
| JM109-1(Master plate) | LB medium
|
"
