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Template:Kyoto/Notebook/Aug 28
From 2013.igem.org
(Difference between revisions)
(→Colony PCR: editor:kojima) |
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===Restriction Enzyme Digestion=== | ===Restriction Enzyme Digestion=== | ||
| + | <div class="experiment"> | ||
| + | <span class="author">No name</span> | ||
| + | {| class="wikitable" | ||
| + | ! ||8/22 RBS-lysis1-(1)||EcoRI||SpeI||BufferB||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||9µL||0.5µL||0.5µL||3µL||0.3µL||16.7µL||30µL | ||
| + | |- | ||
| + | |NC||1µL||0µL||0µL||1µL||0.1µL||7.9µL||10µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/17 RBS-GFP-DT-(2)||XbaI||PstI||BufferD||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 | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/20 Pcon-RBS-GFP-DT-(1)||EcoRI||SpeI||BufferB||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||3µL||0.5µL||0.5µL||3µL||0.3µL||22.7µL||30µL | ||
| + | |- | ||
| + | |NC||0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/20 Pcon-RBS-luxR-DT-(1)||EcoRI||XbaI||BufferD||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||2µL||0.5µL||0.5µL||3µL||0.3µL||23.7µL||30µL | ||
| + | |- | ||
| + | |NC||0.3µL||0µL||0µL||1µL||0.1µL||8.6µL||10µL | ||
| + | |} | ||
| + | </div> | ||
| + | <div class="experiment"> | ||
| + | <span class="author">No name</span> | ||
| + | {| class="wikitable" | ||
| + | ! ||8/28 Plux||SpeI||PstI||BufferD||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||6µL||0.5µL||0.5µL||3µL||0.3µL||19.7µL||30µL | ||
| + | |- | ||
| + | |NC||1µL||0µL||0µL||1µL||0.1µL||7.9µL||10µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/20 RBS-tetR-DT-(1)||XbaI||PstI||BufferD||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||5µL||0.5µL||0.5µL||3µL||0.3µL||20.7µL||30µL | ||
| + | |- | ||
| + | |NC||0.5µL||0µL||0µL||1µL||0.1µL||8.4µL||10µL | ||
| + | |} | ||
| + | </div> | ||
| + | |||
===Electrophoresis=== | ===Electrophoresis=== | ||
===Gel Extraction=== | ===Gel Extraction=== | ||
Revision as of 07:22, 21 September 2013
Contents |
Aug 28
Electrophoresis
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| 8/27 Plux | 173.8 | 1.95 | 1.45 |
Ligation
| state | Vector | Inserter | ||
|---|---|---|---|---|
| experiment | 8/71 DT-1 | 3.0 | 8.0 | |
- Samples were evaporeted used evaporator into about 3 µL.
| sample | MilliQ | Ligation High | total |
|---|---|---|---|
| 3 | 4 | 3.5 | 10.5 |
- incubate one hour at 16 °C
Gel Extraction
| Lane | DNA | |
|---|---|---|
| 1 | 100bpladder | -- |
| 3 | SasA(8/27 Genome PCR production) | |
| 5 | RpaA(8/27 Genome PCR production) | |
| 7 | RpaB(8/27 Genome PCR production) | |
| 9 | PkaiBC(8/27 Genome PCR production) |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Colony PCR
| Sample | base pair |
|---|---|
| 8/21 RBS-lysis2-(3) | 772 |
| 8/21 RBS-lysis2-(4) | 772 |
| 8/21 RBS-lysis2-(5) | 772 |
| 8/21 RBS-lysis2-(6) | 772 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 48s | 30cycles |
Restriction Enzyme Digestion
| 8/22 RBS-lysis1-(1) | EcoRI | SpeI | BufferB | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 9µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 16.7µL | 30µL |
| NC | 1µL | 0µL | 0µL | 1µL | 0.1µL | 7.9µL | 10µL |
| 8/17 RBS-GFP-DT-(2) | XbaI | PstI | BufferD | 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 |
| 8/20 Pcon-RBS-GFP-DT-(1) | EcoRI | SpeI | BufferB | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 3µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 22.7µL | 30µL |
| NC | 0.3µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
| 8/20 Pcon-RBS-luxR-DT-(1) | EcoRI | XbaI | BufferD | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 2µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 23.7µL | 30µL |
| NC | 0.3µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
| 8/28 Plux | SpeI | PstI | BufferD | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 6µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 19.7µL | 30µL |
| NC | 1µL | 0µL | 0µL | 1µL | 0.1µL | 7.9µL | 10µL |
| 8/20 RBS-tetR-DT-(1) | XbaI | PstI | BufferD | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 5µL | 0.5µL | 0.5µL | 3µL | 0.3µL | 20.7µL | 30µL |
| NC | 0.5µL | 0µL | 0µL | 1µL | 0.1µL | 8.4µL | 10µL |
