From 2013.igem.org
Aug 30
No Name
Name | quantity | concentration[µg/mL] | 260/280 | 260/230
|
pT181 attenuator(2) (EcoRI & SpeI) | 305.3 | 2.2 | 1.66 | 0.36
|
pT181 attenuator(2) (XbaI & PstI) | 217.7 | 3.9 | 1.73 | 0.04
|
pT181 antisense(2) (EcoRI & SpeI) | 347.7 | 3.1 | 1.88 | 0.31
|
pT181 antisense(2) (XbaI & PstI) | 376.5 | 5.9 | 0.49 | 2.36
|
pSB1C3 (EcoRI & SpeI) | 436.6 | 30.7 | 1.03 | 1.28
|
pSB1C3 (XbaI & PstI) | 457.4 | 7.9 | 1.65 | 0.42
|
Spinach (EcoRI & SpeI) | 490.5 | 3.0 | 1.87 | 0.26
|
tetR aptamer 12_1R (EcoRI & SpeI) | 451.7 | -12.8 | 5.56 | -0.04
|
Miniprep
No name
DNA | concentration[µg/mL] | 260/280 | 260/230
|
RBS-lysis2 9 | 264.6 | 1.97 | 2.21
|
RBS-lysis2 12 | 220.0 | 1.89 | 2.46
|
RBS-lysis2 14 | 195.3 | 1.88 | 1.53
|
Colony PCR
No name
Sample | base pair
|
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT(8/20 LB Amp) (1)~(4) | 2143
|
8/29 Plux-RBS-GFP-DT(8/27 LB-CP) (1)~(2) | 1227
|
8/29 Pbad/araC-RBS-RFP{BBa_I13516} (8/21 LB-CP) (1)~(2) | 2256
|
8/29 RBS-lysis3-DT-(1) | 1210
|
PreDenature | Denature | Annealing | Extension | cycle
|
94°C | 94°C | 55°C | 68°C | --
|
5min | 30s | 30s | 2min | 30cycle
|
Ethanol Precipitation
No name
DNA
|
pT181 attenuator-(2) (EcoRI & SpeI)
|
pT181 attenuator-(2) (XbaI & PstI)
|
pT181 antisense-(2) (EcoRI & SpeI)
|
pT181 antisense-(2) (XbaI & PstI)
|
pSB1C3 (EcoRI & SpeI)
|
pSB1C3 (XbaI & PstI)
|
Spinach (EcoRI & SpeI)
|
tetR aptamer 12_1R (EcoRI & SpeI)
|
Restriction Enzyme Digestion
No name
| 8/30 RBS-lysis2 (9) | EcoRI | SpeI | 10x bufferB | 100x BSA | MilliQ | total
|
2 cuts | 7.5 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 18.2 µL | 30 µL
|
NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL
|
| 8/22 pSB1C3-(2) | EcoRI | SpeI | 10x Buffer | 100x BSA | MilliQ | total
|
2 cuts | 8 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 17.7 µL | 30 µL
|
NC | 1 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 7.9 µL | 10 µL
|
| 8/22 pSB1C3-(2) | XbaI | PstI | BufferD | BSA | MilliQ | total
|
2 cuts | 8 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 17.7 µL | 30 µL
|
NC | 1 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 7.9 µL | 10 µL
|
| 8/21 Spinach (1) | EcoRI | SpeI | 10x BufferB | 100x BSA | MilliQ | total
|
2 cuts | 4 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 21.9 µL | 30 µL
|
NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL
|
| 8/21 tetR aptamer 12_1R (1) | EcoRI | SpeI | 10xBufferB | 100xBSA | MilliQ | total
|
2 cuts | 3 µL | 0.5 µL | 0.5 µL | 3 µL | 0.3 µL | 22.7 µL | 30 µL
|
NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL
|
| 8/21 pT181 attenuator-(2) | EcoRI | SpeI | XbaI | PstI | 10x BufferB | 10x BufferD | 100x BSA | MilliQ | total
|
2 cuts | 4 µL | 0.5 µL | 0.5 µL | 0 µL | 0 µL | 3 µL | 0 µL | 0.3 µL | 21.7 µL | 30 µL
|
NC | 0.5 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0 µL | 0.1 µL | 8.4 µL | 10 µL
|
2 cuts | 4 µL | 0 µL | 0 µL | 0.5 µL | 0.5 µL | 0 µL | 3 µL | 0.3 µL | 21.7 µL | 30 µL
|
NC | 0.5 µL | 0 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 8.4 µL | 10 µL
|
| 8/21 pT181 antisense-(2) | EcoRI | SpeI | XbaI | PstI | 10x BufferB | 10x BufferD | 100x BSA | MilliQ | total
|
2 cuts | 9 µL | 0.5 µL | 0.5 µL | 0 µL | 0 µL | 3 µL | 0 µL | 0.3 µL | 16.7 µL | 30 µL
|
NC | 1 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0 µL | 0.1 µL | 7.9 µL | 10 µL
|
2 cuts | 9 µL | 0 µL | 0 µL | 0.5 µL | 0.5 µL | 0 µL | 3 µL | 0.3 µL | 16.7 µL | 30 µL
|
NC | 1 µL | 0 µL | 0 µL | 0 µL | 0 µL | 0 µL | 1 µL | 0.1 µL | 7.9 µL | 10 µL
|
Electrophoresis
No name
Lane | Sample | Enzyme1 | Enzyme2
|
1 | 100bp ladder | -- | --
|
2 | RBS-lysis2-(9) | EcoRI | SpeI
|
3 | RBS-lysis2-(9) | -- | --
|
4 | pSB1C3-(2) | EcoRI | SpeI
|
5 | pSB1C3-(2) | -- | --
|
6 | pSB1C3-(2) | XbaI | PstI
|
7 | pSB1C3-(2) | -- | --
|
8 | tRNA-Spinach-(1) | EcoRI | SpeI
|
9 | tRNA-Spinach-(1) | -- | --
|
10 | tetR aptamer12_1R | EcoRI | SpeI
|
11 | tetR aptamer12_1R | -- | --
|
12 | pT181 attenuator | EcoRI | SpeI
|
13 | pT181 attenuator | EcoRI | SpeI
|
14 | 100bp ladder | -- | --
|
File:Igku Aug30electrophoresis1
Lane | Sample | Enzyme1 | Enzyme2
|
1 | 100bp ladder | -- | --
|
2 | pT181 antisense | EcoRI | SpeI
|
3 | pT181 antisense | -- | --
|
4 | pT181 antisense | XbaI | PstI
|
5 | pT181 antisense | -- | --
|
6 | pT181 attenuator | XbaI | PstI
|
7 | pT181 attenuator | -- | --
|
8 | 100bp ladder | -- | --
|
File:Igku Aug30electrophoresis2
Lane | Sample
|
1 | 1kb ladder
|
2 | 8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)(Colony PCR product)
|
3 | 8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)(Colony PCR product)
|
4 | 8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)(Colony PCR product)
|
5 | 8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)(Colony PCR product)
|
6 | Pbad/araC-RBS-RFP-DT-(1)(Colony PCR product)
|
7 | Pbad-araC-RBS-RFP-DT-(2)(Colony PCR product)
|
File:Igku Aug30electrophoresis3