Template:Kyoto/Notebook/Sep 15
From 2013.igem.org
Contents |
Sep 15
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
Pλ-luxI | 225 | 1.59 | 0.83 |
Liquid Culture
Sample | medium |
---|---|
9/13 Pλ-luxI-1 | |
9/7 pSB4K5-1 | |
9/11 Plac-3 |
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | 9/13 Pcon(SpeI & PstI) | 3.6 µL | 9/12 RBS-lacZα-DT(XbaI & PstI) | 4.7 µL | 8 µL |
Restriction Enzyme Digestion
DNA | EcorI | SpeI | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
9/7Pcon-RBS-tetR-DT (EcoRI & SpeI) | 15 µL | 1 µL | 1 µL | 3 µL | 3 µL | 6 µL | 30 µL |
NC | 0.7 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.3 µL | 10 µL |
DNA | EcoRI | SpeI | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
9/12 Pcon-pT181 attenuator-DT (EcoRI & SpeI) | 6.8 µL | 1 µL | 1 µL | 3 µL | 3 µL | 15.2 µL | 30 µL |
NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL |
DNA | EcoRI | SpeI | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
9/12 Pcon-apt12_1R-DT(EcoRI & SpeI) | 6.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 15.4 µL | 30 µL |
NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL |
DNA | EcoRI | SpeI | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
9/8 Pcon-spin-DT (EcoRI & SpeI) | 7.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 15.4 µL | 30 µL |
NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL |
DNA | XbaI | PstI | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
9/11 Pcon-pT181 anticodone-spin-DT(XbaI & PstI) | 7.2 µL | 1 µL | 1 µL | 3 µL | 3 µL | 14.8 µL | 30 µL |
NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.6 µL | 10 µL |
DNA | XbaI | PstI | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
8/17 RBS-lacZα-DT (XbaI & PstI) | 5.3 µL | 1 µL | 1 µL | 3 µL | 3 µL | 16.7 µL | 30 µL |
NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.5 µL | 10 µL |
DNA | EcoRI | XbaI | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
9/7 Ptet-RBS-LacZα-DT (EcoRI & XbaI) | 2.9 µL | 1 µL | 1 µL | 3 µL | 3 µL | 18.5 µL | 30 µL |
NC | 1.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7 µL | 10 µL |
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1,14 | |||
2 | Pcon-RBS-tetR-DT | EcoRI | SpeI |
3 | Pcon-RBS-tetR-DT | ||
4 | Pcon-pT181 attenuator-DT | EcoRI | SpeI |
5 | Pcon-pT181 attenuator-DT | ||
6 | Pcon-apt12-1R-DT | EcoRI | SpeI |
7 | Pcon-apt12-1R-DT | ||
8 | Pcon-spin-DT | EcoRI | SpeI |
9 | Pcon-spin-DT | ||
10 | Pcon-pT181 antisense-spin-DT | XbaI | PstI |
11 | Pcon-pT181 antisense-spin-DT | ||
12 | RBS-lacZα-DT | XbaI | PstI |
13 | RBS-lacZα-DT | ||
15 | Ptet-RBS-lacZα-DT | EcoRI | XbaI |
16 | Ptet-RBS-lacZα-DT | ||
17 | 1kbp ladder |
Gel Extraction
1
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | |
2~4 | Pcon-pT181 attenuator-DT | EcoRI & SpeI |
6~8 | Pcon-apt12_1R-DT | |
10~12 | Pcon-spin-DT |
2
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | |
2~4 | Pcon-pT181 anticodone-spin-DT | XbaI & PstI |
6~8 | RBS-lacZα-DT | XbaI & PstI |
10~12 | Pcon-RBS-tetR-DT | EcoRI & SpeI |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
9/15 Ptet-RBS-lacZα-DT (EcoRI & SpeI) | 1.9 | 1.08 | 0.77 |
9/15 Pcon-RBS-tetR-DT (EcoRI & SpeI) | 5.9 | 1.30 | 0.17 |
9/15 RBS-lacZα-DT (XbaI & PstI) | 1.0 | 0.85 | 0.44 |
Pcon-pT181 anticodone-spin-DT (XbaI & PstI) | 1.3 | 0.95 | 0.28 |
Pcon-spin-DT | 0.9 | 1.37 | 0.05 |
Pcon-apt12_1R-DT | 1.7 | 1.42 | 0.29 |
Pcon-pT181 attenuator-DT (EcoRI & SpeI) | 2.1 | 1.53 | 0.46 |