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Template:Kyoto/Notebook/Sep 2
From 2013.igem.org
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<div class="experiment"> | <div class="experiment"> | ||
<span class="author">Nakamoto</span> | <span class="author">Nakamoto</span> | ||
| + | {| class="wikitable" | ||
| + | ! ||pSB1C3-(1)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts(E+S)||7µL||1µL||1µL||0µL||0µL||3µL||0µL||0.3µL||17.7µL||30µL | ||
| + | |- | ||
| + | |NC(E+S)||0.3µL||0µL||0µL||0µL||0µL||1µL||0µL||0.1µL||8.6µL||10µL | ||
| + | |- | ||
| + | |2 cut(X+P)s||7µL||0µL||0µL||1µL||1µL||0µL||3µL||0.3µL||17.7µL||30µL | ||
| + | |- | ||
| + | |NC(X+P)||0.3µL||0µL||0µL||0µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 tRMA-spinach(1)||EcoRI||SpeI||BufferB||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||8µL||1.0µL||1.0µL||3µL||0.3µL||17.6µL||30µL | ||
| + | |- | ||
| + | |NC||0.5µL||0µL||0µL||1µL||0.1µL||8.4µL||10µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 tetR aptamer12_1R-(1)||EcoRI||SpeI||BufferB||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||6µL||1.0µL||1.0µL||3µL||0.3µL||18.7µL||30µL | ||
| + | |- | ||
| + | |NC||0.4µL||0µL||0µL||1µL||0.1µL||8.5µL||10µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 pT181 attenuator-(2)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts(E+S)||8µL||1µL||1µL||0µL||0µL||3µL||0µL||0.3µL||16.3µL||30µL | ||
| + | |- | ||
| + | |NC(E+S)||0.5µL||0µL||0µL||0µL||0µL||1µL||0µL||0.1µL||8.4µL||10µL | ||
| + | |- | ||
| + | |2 cut(X+P)s||8µL||0µL||0µL||1µL||1µL||0µL||3µL||0.3µL||16.3µL||30µL | ||
| + | |- | ||
| + | |NC(X+P)||0.5µL||0µL||0µL||0µL||0µL||0µL||1µL||0.1µL||8.4µL||10µL | ||
| + | |} | ||
</div> | </div> | ||
Revision as of 06:09, 23 September 2013
Contents |
Sep 2
Liquid Culture
| Sample | medium |
|---|---|
| 9/1 entA-(Master Plate)-1 | 4mL SOB(+Km) |
37°C
Colony PCR
| Sample | base pair |
|---|---|
| 9/1 RBS-lysis2-DT-(1) | 985 |
| Ptet-RBS-lacZα-DT-(1) | 765 |
| Plac-RBS-lacZα-DT-(1) | 765 |
| Plac-RBS-lacZα-DT-(2) | 765 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 1min | 30cycles |
Restriction Enzyme Digestion
| pSB1C3-(1) | EcoRI | SpeI | XbaI | PstI | BufferB | BufferD | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|---|
| 2 cuts(E+S) | 7µL | 1µL | 1µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 17.7µL | 30µL |
| NC(E+S) | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 8.6µL | 10µL |
| 2 cut(X+P)s | 7µL | 0µL | 0µL | 1µL | 1µL | 0µL | 3µL | 0.3µL | 17.7µL | 30µL |
| NC(X+P) | 0.3µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
| 8/21 tRMA-spinach(1) | EcoRI | SpeI | BufferB | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 8µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 17.6µL | 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 | 6µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 18.7µL | 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(E+S) | 8µL | 1µL | 1µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 16.3µL | 30µL |
| NC(E+S) | 0.5µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 8.4µL | 10µL |
| 2 cut(X+P)s | 8µL | 0µL | 0µL | 1µL | 1µL | 0µL | 3µL | 0.3µL | 16.3µL | 30µL |
| NC(X+P) | 0.5µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 8.4µL | 10µL |
Liquid Culture
Master Plate
Gel Extraction
EDTA Solution
Colony PCR
Master Plate
Liquid Culture
Colony PCR
