From 2013.igem.org
Sep 12
Miniprep
Nakamoto
| DNA | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-pT181atte-DT | 293.6 | 1.88 | 1.84
|
| Pcon-apt12-1R-DT | 303.8 | 1.75 | 1.64
|
| Fusion1atte(pSB1c3) | 172.0 | 1.96 | 2.31
|
| RBS-lisis1-DT | 235.1 | 1.59 | 1.60
|
| RBS-lisis2-DT | 387.7 | 1.91 | 2.01
|
| RBS-lisis3-DT | 387.9 | 1.91 | 2.09
|
| Plux | 166.7 | 1.98 | 1.63
|
Electrophoresis
Nakamoto
| Lane | Sample
|
| 1 | 100bp ladder
|
| 2 | 9/11 P-RBS-lisis1-DT(Colony PCR prodution)
|
File:Igku xxxxxx.xxx
Liquid Culture
Nakamoto
| Sample | medium
|
| 9/10 Pcon-apt12-1R-DT1 | Plusgrow medium(+Amp)
|
| 9/10 DT-Pcon-pT181atte1 | Plusgrow medium(+CP)
|
| 9/10 Fusion1-atte(pSB1c3) | Plusgrow medium(+CP)
|
Electrophoresis
Tatsui
1
| Lane | Sample
|
| 1 | 1kb ladder
|
| 2 | pSB4K5(EcoR1&Spe1)
|
| 3 | pSB4K5 NC(EcoR1&Spe1)
|
| 4 | RBS-lacz-DT(EcoR1&Spe1)
|
| 5 | EcoR1&Spe1 NC( EcoR1&Spe1)
|
| 6 | Pcon-RBS-lacz-DT( EcoR1&Spe1)
|
| 7 | Pcon-RBS-lacz-DT NC( EcoR1&Spe1)
|
| 8 | 1kb ladder
|
File:Igku xxxxxx.xxx
2
| Lane | Sample
|
| 1 | 1kb ladder
|
| 2 | Ptet-pT181anti(Spe1&Pst1)
|
| 3 | Ptet-pT181anti NC(Spe1&Pst1)
|
| 4 | RBS-lysis2-DT(Xba1&Pst1)
|
| 5 | EcoR1&Spe1 NC( EcoR1&Spe1)
|
| 6 | --
|
| 7 | --
|
| 8 | 1kb ladder
|
File:Igku xxxxxx.xxx
Tatsui
| Lane | DNA | Enzyme
|
| 1 | 1kb ladder | --
|
| 2 | RBS-lacZ-DT | EcoR1&Spe1
|
| 3 | RBS-lacZ-DT | EcoR1&Spe1
|
| 4 | RBS-lacZ-DT | EcoR1&Spe1
|
| 5 | -- | --
|
| 6 | Pcon-RBS-lacZ-DT | EcoR1&Spe1
|
| 7 | Pcon-RBS-lacZ-DT | EcoR1&Spe1
|
| 8 | Pcon-RBS-lacZ-DT | EcoR1&Spe1
|
| 9 | 1kb ladder | --
|
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| RBS-lacZ-DT(EcoR1&Spe1) | 9.8 | 1.50 | 0.39
|
| Pcon-RBS-lacZ-DT(EcoR1&Spe1) | 11.2 | 1.47 | 0.08
|
Electrophoresis
No name
| Lane | Sample
|
| 1 | 1kb ladder
|
| 2 | Pλ-RBS-luxI-DT(PCR product)
|
| 3 | Pbad/araC-RBS-RFP(PCR product)
|
Colony PCR
Honda
| Sample | base pair
|
| 9/11 apt12-1M(pSB1C3) 1 | 513
|
| 9/11 apt12-1M(pSB1C3) 2 | 513
|
| 9/11 Fusion3m2a09ttenuator(pSB1C3) 1 | 609
|
| 9/11 Fusion6 antisense(pSB1C3)1 | 431-
|
| 9/11 aptamer 12-P(pSB1C3)1 | 515
|
| 9/11 Fusion1 antisense(pSB1C3) 1 | 420
|
| 9/11 Fusion1 antisense(pSB1C3) 2 | 420
|
| 9/11 Plac(BBa-R0011) 1 | 293
|
| 9/11 Plac(BBa-R0011) 2 | 293
|
| 9/11 Plac(BBa-R0011) 3 | 293
|
| 9/11 Plac(BBa-R0011) 4 | 293
|
| 9/11 Pcon-luxR-Plux-GFP 1 | 2138
|
| 9/11 Pcon-RBS-lacZα-DT(pSB4K5) 1 | 712
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 2min12s | 30cycles
|
Restriction Enzyme Digestion
Tatsui
| | 9/5 Pcon | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cuts(P) | 5.7 | 0µL | 0µL | 0µL | 1µL | 3µL | 3µL | 17.3µL | 30µL
|
| NC(P) | 1.9 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.1µL | 10µL
|
| | 8/17 DT | EcoR1 | Buffer | BSA | MilliQ | total
|
| 1cut | 10.6 | 1µL | 3µL | 3µL | 12.4µL | 30µL
|
| NC | 3.5 | 0µL | 1µL | 1µL | 4.5µL | 10µL
|
Colony PCR
Nakamoto,Tatsui
| Sample | base pair
|
| 9/11 Pcon-pT181atte-RBS-lacZα-DT | 965
|
| 9/11 Plux-PBS-lysis1-DT 1 | 613
|
| 9/11 Pcon-pT181attenuator-aptamer12-1R-DT 1 | 859
|
| 9/11 Pcon-pT181attenuator-aptamer12-1R-DT 2 | 859
|
| 9/11 Plux-RBS-lysis3-DT 1 | 1210
|
| 9/11 Plux-RBS-lysis3-DT 2 | 1210
|
| Pcon-Spinach-DT(pSB4K5) 1 | 605
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 1min12s | 30cycles
|
Transformation
tatsui
| Name | Sample | Competent Cells | Plate
|
| 9/7 pSB4K5 | 2µL | 20µL | Amp
|
Electrophoresis
No name
1
| Lane | Sample
|
| 1 | Fusion6 antisense
|
| 2 | Fusion1 antisense-1
|
| 3 | Fusion1 antisense-2
|
| 4 | 100bp ladder
|
| 5 | Plac(BBa-R0011) 1
|
| 6 | Plac(BBa-R0011) 2
|
| 7 | Plac(BBa-R0011) 3
|
| 8 | Plac(BBa-R0011) 4
|
File:Igku xxxxxx.xxx
2
| Lane | Sample
|
| 1 | 1kb ladder
|
| 2 | aptamer12-1M 1
|
| 3 | aptamer12-1M 2
|
| 4 | Fusion3m2 attenuator
|
| 5 | aptamer12-P1
|
| 6 | Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT
|
| 7 | Pcon-RBS-lacZα=DT
|
| 8 | 1kb ladder
|
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)
|
Colony PCR
Nakamoto
| Sample | base pair
|
| 9/11 Fusion1 antisense 3 | 394
|
| 9/11 Fusion1 antisense 4 | 394
|
| 9/11 aptamer12-P 2 | 378
|
| 9/11 aptamer12-P 3 | 378
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 36s | 30cycles
|
File:Igku Aug19electrophoresis1.png
Electrophoresis
Nakamoto
| Lane | Sample | Enzyme
|
| 1 | Fusion1 anti 2 | --
|
| 2 | Fusion1 anti 3 | --
|
| 3 | 1kb ladder | --
|
| 4 | apt12-P 2 | --
|
| 5 | apt12-P 3 | --
|
Electrophoresis
No name
| Lane | Sample | Enzyme
|
| 1 | Pcon-pT181attenuator-RBS-lacZα-DT 1 | --
|
| 2 | Pcon-pT181attenuator-RBS-lacZα-DT 2 | --
|
| 3 | Plux-RBS-lysis1-DT 1 | --
|
| 4 | Pcon-pT181attenuator-aptamer12-1R-DT 1 | --
|
| 5 | Pcon-pT181attenuator-aptamer12-1R-DT 2 | --
|
| 6 | Plux-RBS-lysis3-DT 1 | --
|
| 7 | 1kbp ladder | --
|
| 8 | Plux-RBS-lysis3-DT 2 | --
|
| 9 | Pcon-spinach-DT(pSB4K5) 1
|
| 10 | 9/5 Pcon | Pst1
|
| 11 | 9/5 Pcon NC | --
|
| 12 | 8/17 DT | EcoR1
|
| 13 | 8/17 DT | --
|
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