From 2013.igem.org
Sep 6
Liquid Culture
No name
| Sample | medium
|
| 9/4 RBS-Lysis3＋DT3 | Plusgrow medium(+Cp)
|
| 9/4 RBS-Lysis1＋DT3 | Plusgrow medium(+Cp)
|
| 9/4 PSB4K5 | Plusgrow medium(+K)
|
Colony PCR
No name
| Sample | base pair
|
| 8/27 Pcon-pT181 antisense-(1) | 542
|
| 8/26 pT181 attenuator-DT-(1) | 738
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 30s | 30cycles
|
File:Igku Aug19electrophoresis1.png
Miniprep
No Name
| DNA | concentration [µg/mL] | 260/280 | 260/230
|
| Plac | 153.7 | 2.03 | 2.10
|
| Pλ-luxⅠ-(1) | 76.5 | 2.07 | 2.06
|
| Pλ-luxⅠ-(2) | 86.4 | 2.08 | 2.10
|
| DT | 166.2 | 1.65 | 2.12
|
| J23100 | 190.1 | 1.68 | 2.34
|
Restriction Enzyme Digestion
Tatsui
| | 9/6 RBS-lysis2-DT (78.3 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total
|
| 2 cuts | 22µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 0µL | 30µL
|
| NC | 1.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.7µL | 10µL
|
| | aptamer-DT (251.7 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total
|
| 2 cuts | 0.4µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 14.1µL | 30µL
|
| NC | 0.4µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL
|
| | Pλ-luxI (86.4 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total
|
| 2 cuts | 23.1µL | 1µL | 1µL | 0µL | 0µL | 3µL | 3µL | 1.9µL | 30µL
|
| NC | 1.2µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.8µL | 10µL
|
Colony PCR
No name
| Sample | base pair
|
| 8/27 Pcon-pT181 antisense-(1) | 542
|
| 8/26 pT181 attenuator-DT-(1) | 738
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 30s | 30cycles
|
File:Igku Aug19electrophoresis1.png
No name
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder | --
|
| 2 | 8/20 RBS-lysis1-(1) | EcoRI & SpeI
|
| 3
|
| 4 | -- | --
|
| 5 | 8/28 RBS-GFP-DT-(2) | XbaI & PstI
|
| 6
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Lane | DNA | Enzyme
|
| 1 | 100bp ladder | --
|
| 2 | 8/28 Plux | SpeI & PstI
|
| 3
|
| 4 | -- | --
|
| 5 | 8/20 RBS-tetR-DT-(1) | XbaI & PstI
|
| 6
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-RBS-GFP-DT (EcoRI&SpeI) | 7.1 | 1.88 | 0.07
|
| Pcon-RBS-luxR-DT (EcoRI&XbaI) | 15.3 | 1.93 | 0.70
|
| Plux (SpeI&PstI) | 4.5 | 1.79 | 0.31
|
| RBS-GFP-DT (XbaI&PstI) | 6.0 | 2.16 | 0.03
|
| RBS-tetR-DT (XbaI&PstI) | 7.6 | 1.91 | 0.38
|
Liquid Culture
No name
| Sample | medium
|
| 8/21 RBS-Lysis2-9 | Plusgrow medium(+Amp)
|
| 8/21 RBS-Lysis2-12 | Plusgrow medium(+Amp)
|
| 8/21 RBS-Lysis2-14 | Plusgrow medium(+Amp)
|
Colony PCR
No name
| Sample | base pair
|
| 8/27 Pcon-pT181 antisense-(1) | 542
|
| 8/26 pT181 attenuator-DT-(1) | 738
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 30s | 30cycles
|
File:Igku Aug19electrophoresis1.png
Colony PCR
No name
| Sample | base pair
|
| 8/27 Pcon-pT181 antisense-(1) | 542
|
| 8/26 pT181 attenuator-DT-(1) | 738
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 30s | 30cycles
|
File:Igku Aug19electrophoresis1.png
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