From 2013.igem.org
Sep 13
Miniprep
Nakamoto
| DNA | concentration[µg/mL] | 260/280 | 260/230
|
| 9/12 Fusion6 antisense | 245.9 | 1.88 | 1.41
|
| 9/12 Plux-3 | 115.5 | 1.58 | 1.32
|
| 9/12 Plux-4 | 125.3 | 1.93 | 1.44
|
| 9/12 Fusion3in2 attenuator-1 | 231.9 | 1.81 | 1.31
|
| Lane | DNA | Enzyme
|
| 1 | 1kb ladder | --
|
| 3 | RBS-lysis2-DT | XbaI&PstI
|
| 4 | RBS-lysis2-DT | XbaI&PstI
|
| 5 | RBS-lysis2-DT | XbaI&PstI
|
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39
|
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08
|
</div>
| Lane | DNA | Enzyme
|
| 3 | RBS-lysis3-DT | XbaI&PstI
|
| 4 | RBS-lysis3-DT | XbaI&PstI
|
| 5 | RBS-lysis3-DT | XbaI&PstI
|
| 6 | 1kb ladder | --
|
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39
|
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08
|
</div>
| Lane | DNA | Enzyme
|
| 2 | 1kb ladder | --
|
| 3 | pSB1C3 | EcoRI&SpeI
|
| 4 | pSB1C3 | EcoRI&SpeI
|
| 5 | pSB1C3 | EcoRI&SpeI
|
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39
|
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08
|
</div>
| Lane | DNA | Enzyme
|
| 1 | 1kb ladder | --
|
| 2 | pSB1C3 | XbaI&PstI
|
| 3 | pSB1C3 | XbaI&PstI
|
| 4 | pSB1C3 | XbaI&PstI
|
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39
|
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08
|
</div>
Restriction Enzyme Digestion
Nakamoto
| | 9/13 Plac | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cuts(PstI) | 16 | 0µL | 0µL | 0µL | 1µL | 3µL | 3µL | 7µL | 30µL
|
| NC | 0.8 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.2µL | 10µL
|
| | 9/10 Pcon attenuator | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cut(PstI) | 10 | 0µL | 0µL | 0µL | 1µL | 3µL | 3µL | 13µL | 30µL
|
| NC | 0.5 | 0µL | | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL
|
| | 9/12 Plux | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cuts(PstI) | 12 | 0µL | 0µL | 0µL | 1µL | 3µL | 3µL | 11µL | 30µL
|
| NC | 0.6 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.4µL | 10µL
|
| | 9/12 DT-E | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cut(XbaI) | 23.7 | 0µL | 0µL | 1µL | 0µL | 3µL | 3µL | 0µL | 30.7µL
|
| NC | 1.2 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.8µL | 10µL
|
| | 9/12 Pcon-RBS-luxR-bTE | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cuts(XbaI) | 22.4 | 0µL | 0µL | 1µL | 0µL | 3µL | 3µL | 0.6µL | 30µL
|
| NC | 2.2 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 5.8µL | 10µL
|
| | 9/12 Pcon-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cut(SpeI) | 21 | 0µL | 1µL | 0µL | 0µL | 3µL | 3µL | 12.0µL | 30µL
|
| NC | 2.1 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 5.7µL | 10µL
|
M9 (EDTA)
| volume | 10ml
|
| Na2HPO4 | 300mg
|
| KH2PO4 | 150mg
|
| NaCl | 25mg
|
| NH4Cl | 50mg
|
| Fe(3)-EDTA | 21mg
|
5mM EDTA</div>
| volume | 10ml
|
| Na2HPO4 | 300mg
|
| KH2PO4 | 150mg
|
| NaCl | 25mg
|
| NH4Cl | 50mg
|
| Fe(3)-EDTA | 105mg
|
</div>
Liquid Culture
Tatsui
| Sample | medium
|
| 9/11 aptamer12_1M(pSB1C3)-1 | --
|
| 9/11 aptamer 12_P1 | --
|
| 9/11 Fusaion1 antisense-4 | --
|
| 9/12 pSB4K5-1 | --
|
| 9/12 pSB4K5-2 | --
|
Restriction Enzyme Digestion
Nakamoto
| | 9/13 Plac-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cuts(SpeI) | 23.5 | 0µL | 0.5µL | 0µL | 0µL | 3µL | 3µL | 0µL | 30µL
|
| NC | 8 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 0µL | 10µL
|
| | 9/13 Pcon-pT181 attenuator-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cut(SpeI) | 23.5 | 0µL | 0.5µL | 0µL | 0µL | 3µL | 3µL | 0µL | 30µL
|
| NC | 02.3 | 0µL | | 0µL | 0µL | 0µL | 1µL | 1µL | 5.7µL | 10µL
|
| | 9/13 Plux-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 1cuts(SpeI) | 23.8 | 0µL | 0.2µL | 0µL | 1µL | 3µL | 3µL | 0µL | 30µL
|
| NC | 8 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 0µL | 10µL
|
| | 9/5 RBS-lacZα-DT | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total
|
| 2cut(XbaI&PstI) | 10.8 | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 11.2µL | 30µL
|
| NC | 0.5 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL
|
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