"
Template:Kyoto/Notebook/Sep 6
From 2013.igem.org
Contents |
Sep 6
Liquid Culture
| Sample | medium |
|---|---|
| 9/4 RBS-Lysis3＋DT3 | Plusgrow medium(+Cp) |
| 9/4 RBS-Lysis1＋DT3 | Plusgrow medium(+Cp) |
| 9/4 PSB4K5 | Plusgrow medium(+K) |
Colony PCR
| Sample | base pair |
|---|---|
| Pconst-spinach-DT-(1) | - |
| Pconst-spinach-DT-(2) | - |
| Pconst-spinach-DT-(3) | - |
| Pconst-spinach-DT-(4) | - |
| Plac-spinach-DT-(1) | - |
| Plac-spinach-DT-(2) | - |
| Plac-spinach-DT-(3) | - |
| Plac-spinach-DT-(4) | - |
| Pcon-PT181 antisense-(1) | - |
| Pcon-PT181 antisense-(2) | - |
| Pcon-PT181 antisense-(3) | - |
| Pcon-PT181 antisense-(4) | - |
| Sample | base pair |
|---|---|
| Ptet-PT181 antisense-(1) | - |
| Ptet-PT181 antisense-(2) | - |
| Ptet-PT181 antisense-(3) | - |
| Ptet-PT181 antisense-(4) | - |
| Plac-PT181 attenuater-(1) | - |
| Plac-PT181 attenuater-(2) | - |
| Plac-PT181 attenuater-(3) | - |
| Plac-PT181 attenuater-(4) | - |
Miniprep
| DNA | concentration [µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| Plac | 153.7 | 2.03 | 2.10 |
| Pλ-luxⅠ-(1) | 76.5 | 2.07 | 2.06 |
| Pλ-luxⅠ-(2) | 86.4 | 2.08 | 2.10 |
| DT | 166.2 | 1.65 | 2.12 |
| J23100 | 190.1 | 1.68 | 2.34 |
Restriction Enzyme Digestion
| 9/6 RBS-lysis2-DT (78.3 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 22µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 0µL | 30µL |
| NC | 1.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.7µL | 10µL |
- at 37°C, for 1h
| aptamer-DT (251.7 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 0.4µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 14.1µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
- at 37°C, for 1h
| Pλ-luxI (86.4 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 23.1µL | 1µL | 1µL | 0µL | 0µL | 3µL | 3µL | 1.9µL | 30µL |
| NC | 1.2µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.8µL | 10µL |
- at 37°C, for 1h
Colony PCR
| Sample | base pair |
|---|---|
| 8/27 Pcon-pT181 antisense-(1) | 542 |
| 8/26 pT181 attenuator-DT-(1) | 738 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 30s | 30cycles |
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 2 | 8/20 RBS-lysis1-(1) | EcoRI & SpeI |
| 3 | ||
| 4 | -- | -- |
| 5 | 8/28 RBS-GFP-DT-(2) | XbaI & PstI |
| 6 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | -- |
| 2 | 8/28 Plux | SpeI & PstI |
| 3 | ||
| 4 | -- | -- |
| 5 | 8/20 RBS-tetR-DT-(1) | XbaI & PstI |
| 6 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| Pcon-RBS-GFP-DT (EcoRI&SpeI) | 7.1 | 1.88 | 0.07 |
| Pcon-RBS-luxR-DT (EcoRI&XbaI) | 15.3 | 1.93 | 0.70 |
| Plux (SpeI&PstI) | 4.5 | 1.79 | 0.31 |
| RBS-GFP-DT (XbaI&PstI) | 6.0 | 2.16 | 0.03 |
| RBS-tetR-DT (XbaI&PstI) | 7.6 | 1.91 | 0.38 |
Liquid Culture
| Sample | medium |
|---|---|
| 8/21 RBS-Lysis2-9 | Plusgrow medium(+Amp) |
| 8/21 RBS-Lysis2-12 | Plusgrow medium(+Amp) |
| 8/21 RBS-Lysis2-14 | Plusgrow medium(+Amp) |
Colony PCR
| Sample | base pair |
|---|---|
| 8/27 Pcon-pT181 antisense-(1) | 542 |
| 8/26 pT181 attenuator-DT-(1) | 738 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 30s | 30cycles |
Colony PCR
| Sample | base pair |
|---|---|
| 8/27 Pcon-pT181 antisense-(1) | 542 |
| 8/26 pT181 attenuator-DT-(1) | 738 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 30s | 30cycles |
