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Template:Kyoto/Notebook/Sep 15
From 2013.igem.org
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|experiment||9/13 pSB1C3 XbaI & PstI||1.9 µL||9/15 Pcon-pT181 anticodone-spin-DT XbaI & PstI||4.4 µL||xx µL | |experiment||9/13 pSB1C3 XbaI & PstI||1.9 µL||9/15 Pcon-pT181 anticodone-spin-DT XbaI & PstI||4.4 µL||xx µL | ||
|- | |- | ||
| - | |experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 µL||9/3 pT181 attenuator EcoRI & SpeI|| | + | |experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 µL||9/3 pT181 attenuator EcoRI & SpeI||3.8 µL||xx µL |
|- | |- | ||
| - | |experiment||9/6 Pcon SpeI & PstI||1.4 µL||9/15 RBS-lacZα-DT XbaI & PstI|| | + | |experiment||9/6 Pcon SpeI & PstI||1.4 µL||9/15 RBS-lacZα-DT XbaI & PstI||2.0 µL||xx µL |
|- | |- | ||
| - | |experiment||9/14 pSB 4k5 EcoRI & SpeI|| µL||9/15 Pcon-spin-DT EcoRI & SpeI|| | + | |experiment||9/14 pSB 4k5 EcoRI & SpeI|| µL||9/15 Pcon-spin-DT EcoRI & SpeI||9.4 µL||xx µL |
|- | |- | ||
|experiment||9/15 Peet-RBS-lacZα-DT EcoRI & XbaI||2.6 µL||Pcon-RBS-tetR-DT EcoRI & SpeI||1.5 µL||xx µL | |experiment||9/15 Peet-RBS-lacZα-DT EcoRI & XbaI||2.6 µL||Pcon-RBS-tetR-DT EcoRI & SpeI||1.5 µL||xx µL | ||
Revision as of 15:22, 24 September 2013
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 |
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | 9/13 pSB1C3 EcoRI & SpeI | 4.7 µL | 9/15 Pcon-RBS-tetR-DT EcoRI & SpeI | 1.9 µL | xx µL |
| experiment | 9/13 pSB1C3 EcoRI & SpeI | 4.7 µL | 9/15 Pcon-spin-DT EcoRI & SpeI | 4.7 µL | xx µL |
| experiment | 9/13 pSB1C3 EcoRI & SpeI | 4.7 µL | 9/15 Pcon-apt12_1R-DT EcoRI & SpeI | 2.0 µL | xx µL |
| experiment | 9/13 pSB1C3 EcoRI & SpeI | 4.7 µL | 9/15 Pcon-pT181 attenuator-DT EcoRI & SpeI | 3.0 µL | xx µL |
| experiment | 9/13 pSB1C3 XbaI & PstI | 1.9 µL | 9/15 Pcon-pT181 anticodone-spin-DT XbaI & PstI | 4.4 µL | xx µL |
| experiment | 9/13 pSB1C3 EcoRI & SpeI | 4.7 µL | 9/3 pT181 attenuator EcoRI & SpeI | 3.8 µL | xx µL |
| experiment | 9/6 Pcon SpeI & PstI | 1.4 µL | 9/15 RBS-lacZα-DT XbaI & PstI | 2.0 µL | xx µL |
| experiment | 9/14 pSB 4k5 EcoRI & SpeI | µL | 9/15 Pcon-spin-DT EcoRI & SpeI | 9.4 µL | xx µL |
| experiment | 9/15 Peet-RBS-lacZα-DT EcoRI & XbaI | 2.6 µL | Pcon-RBS-tetR-DT EcoRI & SpeI | 1.5 µL | xx µL |
