Template:Kyoto/Notebook/Sep 22
From 2013.igem.org
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==Sep 22== | ==Sep 22== | ||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC①||283 | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC②||283 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |2min||30s||30s||18s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Miniprep=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {|class="wikitable" | ||
+ | !DNA||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |Pλ-luxI||225 ||1.59 ||0.83 | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC①||283 | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC②||283 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |2min||30s||30s||18s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Gel Extraction=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||DNA||Enzyme | ||
+ | |- | ||
+ | |1||100bp ladder||-- | ||
+ | |- | ||
+ | |3||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |4||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |5||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |6||RBS-lacZα-DT||XbaI&PstI | ||
+ | |} | ||
+ | |||
+ | [[File:igku_Aug19electrophoresis3]]<br> | ||
+ | [[File:igku_Aug19electrophoresis4]]<br> | ||
+ | {| class="wikitable" | ||
+ | !Name||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |RBS-lacZα-DT(XbaI&PstI)||9.8||1.50||0.39 | ||
+ | |- | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===digestion=== | ||
+ | |||
+ | |||
+ | ===Gel Extraction=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||DNA||Enzyme | ||
+ | |- | ||
+ | |1||100bp ladder||-- | ||
+ | |- | ||
+ | |3||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |4||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |5||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |6||RBS-lacZα-DT||XbaI&PstI | ||
+ | |} | ||
+ | |||
+ | [[File:igku_Aug19electrophoresis3]]<br> | ||
+ | [[File:igku_Aug19electrophoresis4]]<br> | ||
+ | {| class="wikitable" | ||
+ | !Name||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |RBS-lacZα-DT(XbaI&PstI)||9.8||1.50||0.39 | ||
+ | |- | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | |||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC①||283 | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC②||283 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |2min||30s||30s||18s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Tatsui</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC①||283 | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC②||283 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |2min||30s||30s||18s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC①||283 | ||
+ | |- | ||
+ | |pSB1C3+P-KaiBC②||283 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |2min||30s||30s||18s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Ligation=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Okazaki, Kanaoka, Tatsui and Hirano</span> | ||
+ | {| class="wikitable" | ||
+ | !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | ||
+ | |- | ||
+ | |experiment||9/18 Plac (SpeI & PstI)||3.0 µL||9/18 RBS-lysis3-DT (XbaI & PstI)||12.4 µL||3.5 µL | ||
+ | |- | ||
+ | |experiment||9/12 Ptet-pT181 antisense||1.0 µL||9/18 spinach-DT||11.9 µL||3.5 µL | ||
+ | |- | ||
+ | |experiment||9/13 Pcon-RBS-luxR-DT||2.1 µL||9/10 Plux-RBS-GFP-DT||5.6 µL||3.5 µL | ||
+ | |- | ||
+ | |experiment||9/13 Pcon-pT181 attenuator (SpeI & PstI)||2.0 µL||9/18 aptamer 12-1R-DT (XbaI & PstI)||6.3 µL||3.5 µL | ||
+ | |- | ||
+ | |experiment||9/18 Plac ②(SpeI & PstI)||3.0 µL||9/18 aptamer 12-1R-DT(XbaI & PstI)||4.5µL||x3.5 µL | ||
+ | |- | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===Liquid Culture=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||medium | ||
+ | |- | ||
+ | |9/2 spinach-DT-2||-- | ||
+ | |- | ||
+ | |8/16 Plux-4||-- | ||
+ | |- | ||
+ | |8/27 Plux-RBS-GFP-DT-1||-- | ||
+ | |- | ||
+ | |9/5 Pcon-pT181 attenuator-1||-- | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===digestion=== | ||
+ | |||
+ | ===Gel Extraction=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||DNA||Enzyme | ||
+ | |- | ||
+ | |1||100bp ladder||-- | ||
+ | |- | ||
+ | |3||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |4||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |5||RBS-lacZα-DT||XbaI&PstI | ||
+ | |- | ||
+ | |6||RBS-lacZα-DT||XbaI&PstI | ||
+ | |} | ||
+ | |||
+ | [[File:igku_Aug19electrophoresis3]]<br> | ||
+ | [[File:igku_Aug19electrophoresis4]]<br> | ||
+ | {| class="wikitable" | ||
+ | !Name||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |RBS-lacZα-DT(XbaI&PstI)||9.8||1.50||0.39 | ||
+ | |- | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | |||
+ | ===Transformation=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Tatsui</span> | ||
+ | {| class="wikitable" | ||
+ | !Name||Sample||Competent Cells||Plate | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-pT181 attenuator-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-aptamer 12_1R-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-antisense-spinach-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-RBS-tetR-DT x2||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/14 pSB1C3-PkaiBC||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/14 pSB1C3-RpaB||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) ||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-pT181 attenuator(XbaI&PstI) ||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-spinach-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/14 RBS-GFP-DT-PkaiBC||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |Plux-RBS-lysis3-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/20 Ptet-pT181 antisense-spinach-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/20 Pcon-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 KaiABC||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 KaiABC||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 Plac-RBS-lysis3-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20 Plac-aptamer 12_1R||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |Plac-aptamer12_1R-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20 Plac-aptamer12_1R-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20Pcon-pT181attenuator-aptamer-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/17 Plac-pT181-attenuatorx2||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/15 Pcon-RBS-lacZα-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/14 Pcon-pT181attenuator-RBS-lacZα-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |PUC19||2 µL||20 µL||Amp | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | ===PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Tatsui</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |pT181 attenuator(pSB1C3)||-- | ||
+ | |- | ||
+ | |Pcon-RBS-lacZα-DT||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94 °C||94 °C||55 °C||68 °C||-- | ||
+ | |- | ||
+ | |5 min||30 s||30 s||3 min||30 cycles | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | |||
+ | ===Transformation=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Tatsui</span> | ||
+ | {| class="wikitable" | ||
+ | !Name||Sample||Competent Cells||Plate | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-pT181 attenuator-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-aptamer 12_1R-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-antisense-spinach-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-RBS-tetR-DT x2||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/14 pSB1C3-PkaiBC||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/14 pSB1C3-RpaB||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) ||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-pT181 attenuator(XbaI&PstI) ||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/17 pSB1C3-Pcon-spinach-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/14 RBS-GFP-DT-PkaiBC||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |Plux-RBS-lysis3-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/20 Ptet-pT181 antisense-spinach-DT||2 µL||20 µL||CP | ||
+ | |- | ||
+ | |9/20 Pcon-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 KaiABC||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 KaiABC||2 µL||20 µL||Kan | ||
+ | |- | ||
+ | |9/20 Plac-RBS-lysis3-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20 Plac-aptamer 12_1R||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |Plac-aptamer12_1R-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20 Plac-aptamer12_1R-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/20Pcon-pT181attenuator-aptamer-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/17 Plac-pT181-attenuatorx2||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/15 Pcon-RBS-lacZα-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |9/14 Pcon-pT181attenuator-RBS-lacZα-DT||2 µL||20 µL||Amp | ||
+ | |- | ||
+ | |PUC19||2 µL||20 µL||Amp | ||
+ | |} | ||
+ | </div> |
Revision as of 15:38, 25 September 2013
Contents |
Sep 22
Colony PCR
Sample | base pair |
---|---|
pSB1C3+P-KaiBC① | 283 |
pSB1C3+P-KaiBC② | 283 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
2min | 30s | 30s | 18s | 30cycles |
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
Pλ-luxI | 225 | 1.59 | 0.83 |
Colony PCR
Sample | base pair |
---|---|
pSB1C3+P-KaiBC① | 283 |
pSB1C3+P-KaiBC② | 283 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
2min | 30s | 30s | 18s | 30cycles |
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | -- |
3 | RBS-lacZα-DT | XbaI&PstI |
4 | RBS-lacZα-DT | XbaI&PstI |
5 | RBS-lacZα-DT | XbaI&PstI |
6 | RBS-lacZα-DT | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
RBS-lacZα-DT(XbaI&PstI) | 9.8 | 1.50 | 0.39 |
digestion
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | -- |
3 | RBS-lacZα-DT | XbaI&PstI |
4 | RBS-lacZα-DT | XbaI&PstI |
5 | RBS-lacZα-DT | XbaI&PstI |
6 | RBS-lacZα-DT | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
RBS-lacZα-DT(XbaI&PstI) | 9.8 | 1.50 | 0.39 |
Colony PCR
Sample | base pair |
---|---|
pSB1C3+P-KaiBC① | 283 |
pSB1C3+P-KaiBC② | 283 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
2min | 30s | 30s | 18s | 30cycles |
Colony PCR
Sample | base pair |
---|---|
pSB1C3+P-KaiBC① | 283 |
pSB1C3+P-KaiBC② | 283 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
2min | 30s | 30s | 18s | 30cycles |
Colony PCR
Sample | base pair |
---|---|
pSB1C3+P-KaiBC① | 283 |
pSB1C3+P-KaiBC② | 283 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
2min | 30s | 30s | 18s | 30cycles |
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | 9/18 Plac (SpeI & PstI) | 3.0 µL | 9/18 RBS-lysis3-DT (XbaI & PstI) | 12.4 µL | 3.5 µL |
experiment | 9/12 Ptet-pT181 antisense | 1.0 µL | 9/18 spinach-DT | 11.9 µL | 3.5 µL |
experiment | 9/13 Pcon-RBS-luxR-DT | 2.1 µL | 9/10 Plux-RBS-GFP-DT | 5.6 µL | 3.5 µL |
experiment | 9/13 Pcon-pT181 attenuator (SpeI & PstI) | 2.0 µL | 9/18 aptamer 12-1R-DT (XbaI & PstI) | 6.3 µL | 3.5 µL |
experiment | 9/18 Plac ②(SpeI & PstI) | 3.0 µL | 9/18 aptamer 12-1R-DT(XbaI & PstI) | 4.5µL | x3.5 µL |
Liquid Culture
Sample | medium |
---|---|
9/2 spinach-DT-2 | -- |
8/16 Plux-4 | -- |
8/27 Plux-RBS-GFP-DT-1 | -- |
9/5 Pcon-pT181 attenuator-1 | -- |
digestion
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | -- |
3 | RBS-lacZα-DT | XbaI&PstI |
4 | RBS-lacZα-DT | XbaI&PstI |
5 | RBS-lacZα-DT | XbaI&PstI |
6 | RBS-lacZα-DT | XbaI&PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
RBS-lacZα-DT(XbaI&PstI) | 9.8 | 1.50 | 0.39 |
Transformation
Name | Sample | Competent Cells | Plate |
---|---|---|---|
9/17 pSB1C3-Pcon-pT181 attenuator-DT | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-aptamer 12_1R-DT | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-antisense-spinach-DT | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-RBS-tetR-DT x2 | 2 µL | 20 µL | CP |
9/14 pSB1C3-PkaiBC | 2 µL | 20 µL | CP |
9/14 pSB1C3-RpaB | 2 µL | 20 µL | CP |
9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) | 2 µL | 20 µL | CP |
9/17 pSB1C3-pT181 attenuator(XbaI&PstI) | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-spinach-DT | 2 µL | 20 µL | CP |
9/14 RBS-GFP-DT-PkaiBC | 2 µL | 20 µL | CP |
Plux-RBS-lysis3-DT | 2 µL | 20 µL | CP |
9/20 Ptet-pT181 antisense-spinach-DT | 2 µL | 20 µL | CP |
9/20 Pcon-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
9/20 KaiABC | 2 µL | 20 µL | Kan |
9/20 KaiABC | 2 µL | 20 µL | Kan |
9/20 Plac-RBS-lysis3-DT | 2 µL | 20 µL | Amp |
9/20 Plac-aptamer 12_1R | 2 µL | 20 µL | Amp |
9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT | 2 µL | 20 µL | Amp |
Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
9/20 Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
9/20Pcon-pT181attenuator-aptamer-DT | 2 µL | 20 µL | Amp |
9/17 Plac-pT181-attenuatorx2 | 2 µL | 20 µL | Amp |
9/15 Pcon-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
9/14 Pcon-pT181attenuator-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
PUC19 | 2 µL | 20 µL | Amp |
PCR
Sample | base pair |
---|---|
pT181 attenuator(pSB1C3) | -- |
Pcon-RBS-lacZα-DT | -- |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94 °C | 94 °C | 55 °C | 68 °C | -- |
5 min | 30 s | 30 s | 3 min | 30 cycles |
Transformation
Name | Sample | Competent Cells | Plate |
---|---|---|---|
9/17 pSB1C3-Pcon-pT181 attenuator-DT | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-aptamer 12_1R-DT | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-antisense-spinach-DT | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-RBS-tetR-DT x2 | 2 µL | 20 µL | CP |
9/14 pSB1C3-PkaiBC | 2 µL | 20 µL | CP |
9/14 pSB1C3-RpaB | 2 µL | 20 µL | CP |
9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) | 2 µL | 20 µL | CP |
9/17 pSB1C3-pT181 attenuator(XbaI&PstI) | 2 µL | 20 µL | CP |
9/17 pSB1C3-Pcon-spinach-DT | 2 µL | 20 µL | CP |
9/14 RBS-GFP-DT-PkaiBC | 2 µL | 20 µL | CP |
Plux-RBS-lysis3-DT | 2 µL | 20 µL | CP |
9/20 Ptet-pT181 antisense-spinach-DT | 2 µL | 20 µL | CP |
9/20 Pcon-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
9/20 KaiABC | 2 µL | 20 µL | Kan |
9/20 KaiABC | 2 µL | 20 µL | Kan |
9/20 Plac-RBS-lysis3-DT | 2 µL | 20 µL | Amp |
9/20 Plac-aptamer 12_1R | 2 µL | 20 µL | Amp |
9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT | 2 µL | 20 µL | Amp |
Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
9/20 Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
9/20Pcon-pT181attenuator-aptamer-DT | 2 µL | 20 µL | Amp |
9/17 Plac-pT181-attenuatorx2 | 2 µL | 20 µL | Amp |
9/15 Pcon-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
9/14 Pcon-pT181attenuator-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
PUC19 | 2 µL | 20 µL | Amp |