Template:Kyoto/Notebook/Aug 29
From 2013.igem.org
(Difference between revisions)
(→Transformation) |
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|Plac||153.7||2.03||2.10 | |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 | |DT||166.2||1.65||2.12 |
Revision as of 12:46, 21 September 2013
Contents |
Aug 29
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 |
Electrophoresis
Lane | Sample |
---|---|
1 | 100bp ladder |
2 | 8/21 RBS-lysis2-9(Colony PCR prodution) |
3 | 8/21 RBS-lysis2-8(Colony PCR prodution) |
4 | 8/21 RBS-lysis2-7(Colony PCR prodution) |
5 | 8/21 RBS-lysis2-10(Colony PCR prodution) |
6 | 8/21 RBS-lysis2-11(Colony PCR prodution) |
7 | 8/21 RBS-lysis2-12(Colony PCR prodution) |
8 | 8/21 RBS-lysis2-13(Colony PCR prodution) |
9 | 8/21 RBS-lysis2-14(Colony PCR prodution) |
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 |
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) |
Restriction Enzyme Digestion
8/22 psB1C3-(2) | EcoRI | SpeI | XbaI | PstI | BufferB | BufferD | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|---|---|---|
2 cuts | 8µL | 0.5µL | 0.5µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 17.7µL | 30µL |
NC | 1µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 7.9µL | 10µL |
2 cuts | 8µL | 0µL | 0µL | 0.5µL | 0.5µL | 0µL | 3µL | 0.3µL | 17.7µL | 30µL |
NC | 1µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 7.9µL | 10µL |
8/21 tRMA-spinach(1) | XbaI | SpeI | BufferB | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 4µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 21.7µL | 30µL |
NC | 0.5µL | 0µL | 0µL | 1µL | 0.1µL | 8.4µL | 10µL |
8/21 tetR aptamer12-1R(1) | EcoRI | SpeI | BufferB | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 3µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 22.7µL | 30µL |
NC | 0.4µL | 0µL | 0µL | 1µL | 0.1µL | 8.5µL | 10µL |
8/21 PT181 attenuator-(2) | EcoRI | SpeI | XbaI | PstI | BufferB | BufferD | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|---|---|---|
2 cuts | 4µL | 0.5µL | 0.5µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 21.7µL | 30µL |
NC | 0.5µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 8.4µL | 10µL |
2 cuts | 4µL | 0µL | 0µL | 0.5µL | 0.5µL | 0µL | 3µL | 0.3µL | 21.7µL | 30µL |
NC | 0.5µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 8.4µL | 10µL |
8/21 PT181 antisense-(2) | EcoRI | SpeI | XbaI | PstI | BufferB | BufferD | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|---|---|---|
2 cuts | 10µL | 0.5µL | 0.5µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 15.7µL | 30µL |
NC | 1µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 7.9µL | 10µL |
2 cuts | 10µL | 0µL | 0µL | 0.5µL | 0.5µL | 0µL | 3µL | 0.3µL | 15.7µL | 30µL |
NC | 1µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 7.9µL | 10µL |
- incubated 37°C 1hour
Transformation
Name | Sample | Competent Cells | Total | Plate |
---|---|---|---|---|
8/27 pT181 attenuator-DT | 2µL | 20µL | 22µL | CP |
8/27 RBS-lysis3-DT | 2µL | 20µL | 22µL | CP |
8/28 RBS-lysis3-DT | 2µL | 20µL | 22µL | CP |
8/28 Pcon-RBS-GFP-DT-Pcon-RBS-LuxR-DT | 2µL | 20µL | 22µL | Amp |
8/28 RBS-lysis1-DT | 2µL | 20µL | 22µL | CP |
8/28 Plux-RBS-GFP-DT | 2µL | 20µL | 22µL | CP |
8/28 Spinach-DT | 2µL | 20µL | 22µL | CP |