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Template:Kyoto/Notebook/Sep 13
From 2013.igem.org
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</div> | </div> | ||
| + | |||
| + | ===Restriction Enzyme Digestion=== | ||
| + | <div class="experiment"> | ||
| + | <span class="author">Nakamoto</span> | ||
| + | {| class="wikitable" | ||
| + | ! ||9/13 Plac-P||EcoRI||SpeI||XbaI||PstI|| Buffer||BSA||MilliQ||total | ||
| + | |- | ||
| + | |1cuts(SpeI)||23.5||0µL||0.5µL||0µL||0µL||3µL||3µL||0µL||30µL | ||
| + | |- | ||
| + | |NC||8||0µL||0µL||0µL||0µL||1µL||1µL||0µL||10µL | ||
| + | |- | ||
| + | |} | ||
| + | |||
| + | {| class="wikitable" | ||
| + | ! ||9/13 Pcon-pT181 attenuator-P||EcoRI||SpeI||XbaI||PstI||Buffer||BSA||MilliQ||total | ||
| + | |- | ||
| + | |1cut(SpeI)||23.5||0µL||0.5µL||0µL||0µL||3µL||3µL||0µL||30µL | ||
| + | |- | ||
| + | |NC||02.3||0µL||||0µL||0µL||0µL||1µL||1µL||5.7µL||10µL | ||
| + | |} | ||
| + | |||
| + | {| class="wikitable" | ||
| + | ! ||9/13 Plux-P ||EcoRI||SpeI||XbaI||PstI|| Buffer||BSA||MilliQ||total | ||
| + | |- | ||
| + | |1cuts(SpeI)||23.8||0µL||0.2µL||0µL||1µL||3µL||3µL||0µL||30µL | ||
| + | |- | ||
| + | |NC||8||0µL||0µL||0µL||0µL||1µL||1µL||0µL||10µL | ||
| + | |- | ||
| + | |} | ||
| + | |||
| + | {| class="wikitable" | ||
| + | ! ||9/5 RBS-lacZα-DT||EcoRI||SpeI||XbaI||PstI||Buffer||BSA||MilliQ||total | ||
| + | |- | ||
| + | |2cut(XbaI&PstI)||10.8||0µL||0µL||1µL||1µL||3µL||3µL||11.2µL||30µL | ||
| + | |- | ||
| + | |NC||0.5||0µL||0µL||0µL||0µL||1µL||1µL||7.5µL||10µL | ||
| + | |} | ||
Revision as of 07:53, 25 September 2013
Contents |
Sep 13
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| 9/12 Fusion6 antisense | 245.9 | 1.88 | 1.41 |
| 9/12 Plux-3 | 115.5 | 1.58 | 1.32 |
| 9/12 Plux-4 | 125.3 | 1.93 | 1.44 |
| 9/12 Fusion3in2 attenuator-1 | 231.9 | 1.81 | 1.31 |
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kb ladder | -- |
| 2 | RBS-lysis1-DT | XbaI&PstI |
| 3 | RBS-lysis1-DT | XbaI&PstI |
| 4 | RBS-lysis1-DT | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39 |
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08 |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kb ladder | -- |
| 3 | RBS-lysis2-DT | XbaI&PstI |
| 4 | RBS-lysis2-DT | XbaI&PstI |
| 5 | RBS-lysis2-DT | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39 |
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08 |
</div>
| Lane | DNA | Enzyme |
|---|---|---|
| 3 | RBS-lysis3-DT | XbaI&PstI |
| 4 | RBS-lysis3-DT | XbaI&PstI |
| 5 | RBS-lysis3-DT | XbaI&PstI |
| 6 | 1kb ladder | -- |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39 |
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08 |
</div>
| Lane | DNA | Enzyme |
|---|---|---|
| 2 | 1kb ladder | -- |
| 3 | pSB1C3 | EcoRI&SpeI |
| 4 | pSB1C3 | EcoRI&SpeI |
| 5 | pSB1C3 | EcoRI&SpeI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39 |
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08 |
</div>
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kb ladder | -- |
| 2 | pSB1C3 | XbaI&PstI |
| 3 | pSB1C3 | XbaI&PstI |
| 4 | pSB1C3 | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39 |
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08 |
</div>
Restriction Enzyme Digestion
| 9/13 Plac | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 1cuts(PstI) | 16 | 0µL | 0µL | 0µL | 1µL | 3µL | 3µL | 7µL | 30µL |
| NC | 0.8 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.2µL | 10µL |
| 9/10 Pcon attenuator | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | ||
|---|---|---|---|---|---|---|---|---|---|---|
| 1cut(PstI) | 10 | 0µL | 0µL | 0µL | 1µL | 3µL | 3µL | 13µL | 30µL | |
| NC | 0.5 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL |
| 9/12 Plux | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 1cuts(PstI) | 12 | 0µL | 0µL | 0µL | 1µL | 3µL | 3µL | 11µL | 30µL |
| NC | 0.6 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.4µL | 10µL |
| 9/12 DT-E | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 1cut(XbaI) | 23.7 | 0µL | 0µL | 1µL | 0µL | 3µL | 3µL | 0µL | 30.7µL |
| NC | 1.2 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.8µL | 10µL |
| 9/12 Pcon-RBS-luxR-bTE | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 1cuts(XbaI) | 22.4 | 0µL | 0µL | 1µL | 0µL | 3µL | 3µL | 0.6µL | 30µL |
| NC | 2.2 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 5.8µL | 10µL |
| 9/12 Pcon-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 1cut(SpeI) | 21 | 0µL | 1µL | 0µL | 0µL | 3µL | 3µL | 12.0µL | 30µL |
| NC | 2.1 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 5.7µL | 10µL |
M9 (EDTA)
1mM EDTA
| volume | 10ml |
|---|---|
| Na2HPO4 | 300mg |
| KH2PO4 | 150mg |
| NaCl | 25mg |
| NH4Cl | 50mg |
| Fe(3)-EDTA | 21mg |
5mM EDTA</div>
| volume | 10ml |
|---|---|
| Na2HPO4 | 300mg |
| KH2PO4 | 150mg |
| NaCl | 25mg |
| NH4Cl | 50mg |
| Fe(3)-EDTA | 105mg |
</div>
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kb ladder | -- |
| 3 | Pcon-P | SpeI |
| 4 | Pcon-P | SpeI |
| 5 | Pcon-P | SpeI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lacZα-DT(EcoRI&SpeI) | 9.8 | 1.50 | 0.39 |
| Pcon-RBS-lacZα-DT(EcoRI&SpeI) | 11.2 | 1.47 | 0.08 |
Liquid Culture
| Sample | medium |
|---|---|
| 9/11 aptamer12_1M(pSB1C3)-1 | -- |
| 9/11 aptamer 12_P1 | -- |
| 9/11 Fusaion1 antisense-4 | -- |
| 9/12 pSB4K5-1 | -- |
| 9/12 pSB4K5-2 | -- |
Restriction Enzyme Digestion
| 9/13 Plac-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 1cuts(SpeI) | 23.5 | 0µL | 0.5µL | 0µL | 0µL | 3µL | 3µL | 0µL | 30µL |
| NC | 8 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 0µL | 10µL |
| 9/13 Pcon-pT181 attenuator-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | ||
|---|---|---|---|---|---|---|---|---|---|---|
| 1cut(SpeI) | 23.5 | 0µL | 0.5µL | 0µL | 0µL | 3µL | 3µL | 0µL | 30µL | |
| NC | 02.3 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 5.7µL | 10µL |
| 9/13 Plux-P | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 1cuts(SpeI) | 23.8 | 0µL | 0.2µL | 0µL | 1µL | 3µL | 3µL | 0µL | 30µL |
| NC | 8 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 0µL | 10µL |
| 9/5 RBS-lacZα-DT | EcoRI | SpeI | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2cut(XbaI&PstI) | 10.8 | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 11.2µL | 30µL |
| NC | 0.5 | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.5µL | 10µL |
