From 2013.igem.org
(Difference between revisions)
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| | </div> | | </div> |
| | <!-- ここまでをコピーしてね --> | | <!-- ここまでをコピーしてね --> |
| - | ===Electrophoresis===
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| - | <div class="experiment">
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| - | <span class="author">Okazaki</span>
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| - | {| class="wikitable"
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| - | !Lane||Sample||Enzyme1||Enzyme2
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| - | |-
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| - | |1||1kbpladder||--||--
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| - | |-
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| - | |2||Pcon-PT181antisense-spinach-DT||XbaI||PstI
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| - | |-
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| - | |3||spinach-DT||XbaI||PstI
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| - | |-
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| - | |4||RBS-lacZα-DT||XbaI||PstI
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| - | |-
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| - | |5||Pcon-PT181-antisense-spinach-DT||XbaI||PstI
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| - | |-
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| - | |6||aptamer||XbaI||PstI
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| - | |-
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| - | |7||PT181-attenuator-21:00~||XbaI||PstI
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| - | |-
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| - | |8||Pcon-attenuator||EcoRI||SpeI
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| - | |}
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| - | [[File:igku_xxxxxx.xxx]]<br>
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| - | </div>
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| - | <!-- ここまでをコピーしてね -->
| |
| - | <div class="experiment">
| |
| - | <span class="author">Okazaki</span>
| |
| - | {| class="wikitable"
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| - | !Lane||Sample||Enzyme1||Enzyme2
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| - | |-
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| - | |1||1kbpladder||--||--
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| - | |-
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| - | |2||Pcon-PT181antisense-spinach-DT||--||--
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| - | |-
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| - | |3||spinach-DT||--||--
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| - | |-
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| - | |4||RBS-lacZα-DT||--||--
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| - | |-
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| - | |5||Pcon-PT181-antisense-spinach-DT||--||--
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| - | |-
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| - | |6||aptamer||--||--
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| - | |-
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| - | |7||PT181-attenuator-21:00~||--||--
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| - | |-
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| - | |8||Pcon-attenuator||--||--
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| - | |}
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| - | [[File:igku_xxxxxx.xxx]]<br>
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| - | </div>
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| - |
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| | ===Ligation=== | | ===Ligation=== |
| | | | |
Revision as of 08:52, 24 September 2013
Sep 17
Liquid Culture
Tatsui and Hirano
| Sample | medium
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| Spi-DT | LB medium(+CP)
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| Pcon-Spi-DT,Pcon-pT181-anti-spi-DT | LB medium(+Amp)
|
PCR
Hirano
| Pcon-tet ap-DT(VF) | BigDye | 5x buffer | primer(VF) | MilliQ | total
|
| 1.3 | 0.5 | 1.75 | 0.5 | 0.45 | 10.5
|
| PreDenature | Denature | Annealing | Extension | cycle
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| 96°C | 96°C | 50°C | 60°C | --
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| 2m | 10s | 5s | 4m | 30cycles
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| Pcon-tet ap-DT(VR) | BigDye | 5x buffer | primer(VR) | MilliQ | total
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| 1.3 | 0.5 | 1.75 | 0.5 | 0.45 | 10.5
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| PreDenature | Denature | Annealing | Extension | cycle
|
| 96°C | 96°C | 50°C | 60°C | --
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| 2m | 10s | 5s | 4m | 30cycles
|
Electrophoresis
No name
| Lane | Sample | Enzyme1 | Enzyme2
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| 1 | 1kbpladder | -- | --
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| 2 | Pcon-RBS-tetR-DT | XbaI | PstI
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| 3 | Pcon-RBS-tetR-DT | -- | --
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| 4 | pSB4K5X7 | EcoRI | XbaI
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| 5 | pSB4K5X7 | -- | --
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| 6 | PSB1C3 | EcoRI | SpeI
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| 7 | PSB1C3 | EcoRI | PstI
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| 8 | PSB1C3 | -- | --
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| 9 | pT181attenuator | -- | --
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| 10 | pT181attenuator | -- | --
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| 11 | 1kbpladder | -- | --
|
File:Igku xxxxxx.xxx
Ligation
No Name
| state | Vector | Inserter | Ligation High ver.2
|
| experiment | 8/28 Plux(SpeI+PstI) | 45µg/mL | 9/13 RBS-lysis1-DT (XbaI+PstI) | 7.8µg/mL | xx µg/mL
|
| experiment | 8/28 Plux(SpeI+PstI) | 45µg/mL | 9/13 RBS-lysis2-DT (XbaI+PstI) | 11.8µg/mL | xx µg/mL
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| experiment | 8/28 Plux(SpeI+PstI) | 45µg/mL | 9/13 RBS-lysis3-DT (XbaI+PstI) | 11.8µg/mL | xx µg/mL
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| experiment | 9/14 Plac(SpeI+PstI) | 21µg/mL | 9/16 PT181 atte-DT (XbaI+PstI) | 8.2µg/mL | xx µg/mL
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| experiment | 9/14 Plac(SpeI+PstI) | 21µg/mL | 9/3 PT181 anti (XbaI+PstI) | 6.6µg/mL | xx µg/mL
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| experiment | 9/14 Plac(SpeI+PstI) | 21µg/mL | 9/10 apt12-1R-DT(XbaI+PstI) | 15.6µg/mL | xx µg/mL
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| experiment | 9/13 pSB1C3(XbaI+PstI) | 26.0µg/mL | 9/15 Pcon-PT181 anti-spin-DT(XbaI+PstI) | 5.0µg/mL | xx µg/mL
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| experiment | 9/16 pSB1C3(EcoRI+SpeI) | 49.5µg/mL | 9/15 Pcon-spin-DT(EcoRI+SpeI) | 9.0µg/mL | xx µg/mL
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| experiment | 9/16 pSB1C3(EcoRI+SpeI) | 49.5µg/mL | 9/15 Pcon-RBS-tetR-DT(EcoRI+SpeI) | 59.0µg/mL | xx µg/mL
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| experiment | 9/16 pSB1C3(EcoRI+SpeI) | 49.5µg/mL | 9/15 Pcon-PT181-atte-DT(EcoRI+SpeI) | 21.0µg/mL | xx µg/mL
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| experiment | 9/16 pSB1C3(EcoRI+SpeI) | 49.5µg/mL | 9/15 Pcon-apt12-112-DT(EcoRI+SpeI) | 17.0µg/mL | xx µg/mL
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| Lane | DNA | Enzyme
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| 1 | 1kbpladder | --
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| 2 | pSB1C3 | EcoI+SpeI
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| 3
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| 4
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File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
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| pSB1C3 | 10.0 | 1.84 | 0.40
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| Lane | DNA | Enzyme
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| 1 | 1kbpladder | --
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| 2 | pSB4K5 | XbaI+PstI
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| 3
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| 4
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File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
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| pSB4K5 | 7.6 | 1.81 | 0.32
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| Lane | DNA | Enzyme
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| 1 | 1kbpladder | --
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| 2 | Pcon-RBS-tetR-DT | XbaI+PstI
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| 3
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| 4
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File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
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| Pcon-RBS-tetR-DT | 8.0 | 2.40 | 1.14
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| Lane | DNA | Enzyme
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| 1 | 1kbpladder | --
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| 2 | pSB1C3 | EcoRI+PstI
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| 3
|
| 4
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File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
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| pSB1C3 | 9.1 | 2.17 | 0.35
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Mutation PCR
Kinose
| pC KaiABC(100pg) | Prime STAR(2x) | Primer(KaiA-Bsai MuT-Fwd) | Primer(KaiA-Bsai MuT-Rev) | MilliQ | total
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| 1.0 | 25.0 | 1.0 | 1.0 | 22.0 | 50.0
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| PreDenature | Denature | Annealing | Extension | cycle
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| 96°C | 98°C | 65°C | 72°C | --
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| 5m | 10s | 15s | 40s | 30cycles
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| pC KaiABC(150pg) | Prime STAR(2x) | Primer(KaiA-Bsai MuT-Fwd) | Primer(KaiA-Bsai MuT-Rev) | MilliQ | total
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| 1.5 | 25.0 | 1.0 | 1.0 | 21.5 | 50.0
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| PreDenature | Denature | Annealing | Extension | cycle
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| 96°C | 98°C | 65°C | 72°C | --
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| 5m | 10s | 15s | 40s | 30cycles
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| pC KaiABC(200pg) | Prime STAR(2x) | Primer(KaiA-Bsai MuT-Fwd) | Primer(KaiA-Bsai MuT-Rev) | MilliQ | total
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| 2 | 25.0 | 1.0 | 1.0 | 21.0 | 50.0
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| PreDenature | Denature | Annealing | Extension | cycle
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| 96°C | 98°C | 65°C | 72°C | --
|
| 5m | 10s | 15s | 40s | 30cycles
|
"
