Template:Kyoto/Notebook/Sep 6
From 2013.igem.org
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!Sample||medium | !Sample||medium | ||
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- | | | + | |9/4 RBS-Lysis3+DT3||Plusgrow medium(+Cp) |
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- | | | + | |9/4 RBS-Lysis1+DT3||Plusgrow medium(+Cp) |
|- | |- | ||
- | | | + | |9/4 PSB4K5||Plusgrow medium(+K) |
|} | |} | ||
</div> | </div> |
Revision as of 09:28, 24 September 2013
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 |
---|---|
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 |
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 |