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Template:Kyoto/Notebook/Sep 7
From 2013.igem.org
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Revision as of 13:34, 24 September 2013
Contents |
Sep 7
Gel Extraction
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| apt 12_1R-DT | 12.7 | 1.62 | 5.74 |
| RBS-lysis2-DT | 6.7 | 1.71 | 5.46 |
| Pλ-luvI | 8.1 | 1.76 | 2.25 |
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lysis1-DT | 195.3 | ||
| RBS-lysis3-DT | 255.5 | 1.91 | 2.04 |
| Ptet-pT18 | 183.6 | 1.85 | 1.48 |
| Pcon-RBS-tetR-DT | 134.3 | 1.94 | 2.20 |
| Ptet-RBS-lacZα-DT | 67.7 | 1.92 | 1.79 |
| Pcon-pT181 attenuator | 232.3 | 1.95 | 1.90 |
| pSB4K5 | 237.0 | 1.61 | 1.50 |
| pT181 antisense(pSB1C3) | 256.4 | 1.63 | 1.78 |
| apt12_1R(pSB1C3) | 230.0 | 1.98 | 2.28 |
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | 9/6 Plux (SpeI&PstI) | 2 µL | 9/7 RBS-lysis2-DT (EcoRI&XbaI) | µL | 3.5 µL |
| experiment | 9/6 Plac (SpeI&PstI) | µL | 9/7 RBS-lysis2-DT (EcoRI&XbaI) | µL | 3.5 µL |
| experiment | 9/6 Pcon-RBS-luxR-DT (EcoRI&XbaI) | 1.9 µL | 9/6 Plux-RBS-GFP-DT(SpeI&PstI) | 2.2 µL | 3.5 µL |
| experiment | 9/6 Pbed/araC (SpeI&PstI) | 1.6 µL | 9/5 RBS-luxI-DT (EcoRI&XbaI) | 16 µL | 3.5 µL |
| experiment | 9/6 Plac (SpeI&PstI) | 1.5 µL | 9/5 pT181 antisense (EcoRI&XbaI) | 2.0 µL | 3.5 µL |
| experiment | 9/6 Pcon (SpeI&PstI) | 3.0 µL | 9/5 RBS-luxI-DT (EcoRI&XbaI) | 10 µL | 3.5 µL |
Colony PCR
| Sample | base pair |
|---|---|
| 9/5 Plac-spinarch-DT-13~20 | |
| 9/5 Plac-pT181 attenuator-13~20 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94 °C | 94 °C | 55 °C | 60 °C | -- |
| 5min | 30s | 30s | 42s | 30cycle |
Transformation
| Name | Sample | Competent Cells | Total | Plate |
|---|---|---|---|---|
| Pcon-RBS-luxR-DT+Plux-RBS-GFP-DT | 2µL | 20µL | 22µL | |
| Pbed/araC+RBS-luxI-DT | 2µL | 20µL | 22µL | |
| Plac+pT181 antisence | 2µL | 20µL | 22µL | |
| Pcon+RBS-luxI-DT | 2µL | 20µL | 22µL |
Restriction Enzyme Digestion
| DNA | XbaI | PstI | Buffer | BSA | MilliQ | total | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 9/7 RBS-lysis1-DT(XbaI & PstI) | 10 µL | 1 µL | 1 µL | 3 µL | 3 µL | 12 µL | 30 µL | ||||||||
| NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.5 µL | 10 µL | DNA | XbaI | PstI | Buffer | BSA | MilliQ | total | |
| 9/7 RBS-lysis3-DT (XbaI & PstI) | 7.8 µL | 1 µL | 1 µL | 3 µL | 3 µL | 14.2 µL | 30 µL | ||||||||
| NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.6 µL | 10 µL
| DNA | SpeI | PstI | Buffer | BSA | MilliQ | total | |
| 9/5 Plac (SpeI & PstI) | 6.0 µL | 1 µL | 1 µL | 3 µL | 3 µL | 16.0 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 | |
| 9/7 pSB4K5 (EcoRI & SpeI) | 8.4 µL | 1 µL | 1 µL | 3 µL | 3 µL | 16.0 µL | 30 µL | ||||||||
| NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL
| DNA | restricion enzyme | restriction enzyme | Buffer | BSA | MilliQ | total | |
| Pcon-pT181 attenuator (SpeI & PstI) | 8.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 13.4 µL | 30 µL | ||||||||
| Pcon-pT181 attenuator (EcorI & SpeI) | 8.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 13.4 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 | |
| 9/5 Spin-DT (XbaI & PstI) | 5.4 µL | 1 µL | 1 µL | 3 µL | 3 µL | 16.6 µL | 30 µL | ||||||||
| NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL
| DNA | SpeI | PstI | Buffer | BSA | MilliQ | total | |
| Ptet-anticodone (SpeI & PstI) | 11 µL | 1 µL | 1 µL | 3 µL | 3 µL | 11 µL | 30 µL | ||||||||
| NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.5 µL | 10 µL
| DNA | XbaI | PstI | Buffer | BSA | MilliQ | total | |
| 9/6 RBS-lacZα-DT (XbaI & PstI) | 11 µL | 1 µL | 1 µL | 3 µL | 3 µL | 11 µL | 30 µL | ||||||||
| NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.5 µL | 10 µL
| DNA | EcoRI | SpeI | Buffer | BSA | MilliQ | total | |
| pSB1C3 (1) (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 | XbaI | PstI | Buffer | BSA | MilliQ | total | |
| pSB1C3 (3) (XbaI & PstI) | 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 |
