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Template:Kyoto/Notebook/Sep 3
From 2013.igem.org
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!Plac(1) plasmid 153.7ng/µL||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer (KaiA_BsaI_Mut_fwd)||primer (KaiA_BsaI_Mut_rer||KOD plus-ver.2||MilliQ||total | !Plac(1) plasmid 153.7ng/µL||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer (KaiA_BsaI_Mut_fwd)||primer (KaiA_BsaI_Mut_rer||KOD plus-ver.2||MilliQ||total | ||
|- | |- | ||
| - | |0.13 ||2.5||2.5||1.5||0.75||0.75||0.5||16.37||25|| | + | |0.13 ||2.5||2.5||1.5||0.75||0.75||0.5||16.37||25 |
| + | |} | ||
| + | </div> | ||
| + | <div class="experiment"> | ||
| + | {| class="wikitable" | ||
| + | !KaiB (plasmid) 20.2ng/µL||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer (GG2_RBS_KaiB_fwd)||primer (KaiB_GG3_rer||KOD plus-ver.2||MilliQ||total | ||
| + | |- | ||
| + | |1||2.5||2.5||1.5||0.75||0.75||0.5||15.5||25 | ||
|} | |} | ||
{| class="wikitable" | {| class="wikitable" | ||
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|} | |} | ||
</div> | </div> | ||
| + | |||
<!-- ここまでをコピーしてね --> | <!-- ここまでをコピーしてね --> | ||
Revision as of 12:16, 25 September 2013
Contents |
Sep 3
File:Igku Notebook Sep3 1.jpg File:Igku Notebook Sep3 2.jpg File:Igku Notebook Sep3 3 1.jpg File:Igku Notebook Sep3 3 2.jpg File:Igku Notebook Sep3 4 1.jpg File:Igku Notebook Sep3 4 2.jpg File:Igku Notebook Sep3 4 3.jpg File:Igku Notebook Sep3 4 4.jpg File:Igku Notebook Sep3 5 1.jpg File:Igku Notebook Sep3 5 2.jpg File:Igku Notebook Sep3 5 3.jpg File:Igku Notebook Sep3 5 4.jpg
Electrophoresis
| Lane | Sample |
|---|---|
| 1 | 100bp ladder |
| 2 | pT181 attenuater(2)-DT2(738) |
| 3 | Ptet-RBS-lacZα-DT-2(756) |
| 4 | Ptet-RBS-lacZα-DT-3 |
| 5 | Ptet-RBS-lacZα-DT-4 |
| 6 | Ptet-RBS-lacZα-DT-5 |
| 7 | Ptet-RBS-lacZα-DT-6 |
| 8 | RBS-lysis2-DT-2(985) |
| 9 | RBS-lysis2-DT-3 |
| 10 | 100bp ladder |
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Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| 9/2 14:00 pT181 attenuator | 418.4 | 1.82 | 1.80 |
| 9/2 14:00 pT181 antisense | 362.2 | 1.92 | 1.96 |
| 9/2 14:00 Spinach | 438.3 | 1.86 | 1.99 |
| 9/2 14:00 tetR-apt 12_1R | 567.5 | 1.90 | 1.99 |
| 9/2 21:00 pT181 attenuater | 320.9 | 1.90 | 2.15 |
| 9/2 21:00 Spinach | 457.0 | 1.88 | 2.24 |
Liquid Culture
| Sample | medium |
|---|---|
| Spinach-DT(9/2 master plate) | plusgrow(CP)4ml |
Restriction Enzyme Digestion
| DNA | EcoR1 | Spe1 | Xba1 | Pst1 | 10xBuffer B(EcoR1&Spe1) | 10xBuffer D(Xba1&Pst1) | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|---|
| 9/3 pT181attenuater 14:00 418.4ng/µL | 4.8µL | 1.0µL | 1.0µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 19.9µL | 30µL |
| 9/3 pT181attenuater 14:00 418.4ng/µL | 0.2µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 8.7µL | 10µL |
| 9/3 pT181attenuater 14:00 418.4ng/µL | 4.8µL | 0µL | 0µL | 1.0µL | 1.0µL | 0µL | 3µL | 0.3µL | 19.9µL | 30µL |
| 9/3 pT181attenuater 14:00 418.4ng/µL | 0.2µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 8.7µL | 10µL |
| DNA | EcoR1 | Spe1 | Xba1 | Pst1 | 10xBuffer B(EcoR1&Spe1) | 10xBuffer D(Xba1&Pst1) | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|---|
| 9/3 pT181antisense 14:00 362.2ng/µL | 5.5µL | 1.0µL | 1.0µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 19.2µL | 30µL |
| 9/3 pT181attenuater 14:00 418.4ng/µL | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 8.6µL | 10µL |
| 9/3 pT181attenuater 14:00 418.4ng/µL | 5.5µL | 0µL | 0µL | 1.0µL | 1.0µL | 0µL | 3µL | 0.3µL | 19.2µL | 30µL |
| 9/3 pT181attenuater 14:00 418.4ng/µL | 0.3µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
| DNA | EcoR1 | Spe1 | 10xBuffer B | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 9/3 apt 12_1R 14:00 507.5ng/µL | 3.9µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 20.8µL | 30µL |
| 9/3 apt 12_1R 14:00 507.5ng/µL | 0.2µL | 0µL | 0µL | 1µL | 0.1µL | 8.7µL | 10µL |
| DNA | EcoR1 | Xba1 | 10xBuffer D | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 8/29 DT 166.2ng/µL | 12µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 12.7µL | 30µL |
| 8/29 DT 166.2ng/µL | 0.6µL | 0µL | 0µL | 1µL | 0.1µL | 8.3µL | 10µL |
| DNA | EcoR1 | Spe1 | 10xBuffer B | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 8/22 RBS-lysis1(2) 96ng/µL | 21µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 3.7µL | 30µL |
| 8/22 RBS-lysis1(2) 96ng/µL | 1.0µL | 0µL | 0µL | 1µL | 0.1µL | 7.9µL | 10µL |
| DNA | EcoR1 | Spe1 | 10xBuffer B | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 8/30 RBS-lysis2 220ng/µL | 9.1µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 15.6µL | 30µL |
| 8/30 RBS-lysis2 220ng/µL | 0.5µL | 0µL | 0µL | 1µL | 0.1µL | 8.4µL | 10µL |
| DNA | EcoR1 | Spe1 | 10xBuffer B | 100xBSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 8/20 RBS-lysis3(1) 282ng/µL | 7.1µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 17.6µL | 30µL |
| 8/20 RBS-lysis3(1) 282ng/µL | 0.4µL | 0µL | 0µL | 1µL | 0.1µL | 8.5µL | 10µL |
PCR
| Plac(1) plasmid 153.7ng/µL | 10x buffer KOD plus-ver.2 | 2mM dNTPs | 25mM MgSO4 | primer (KaiA_BsaI_Mut_fwd) | primer (KaiA_BsaI_Mut_rer | KOD plus-ver.2 | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 0.13 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 16.37 | 25 |
| KaiB (plasmid) 20.2ng/µL | 10x buffer KOD plus-ver.2 | 2mM dNTPs | 25mM MgSO4 | primer (GG2_RBS_KaiB_fwd) | primer (KaiB_GG3_rer | KOD plus-ver.2 | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 1 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.5 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| °C | °C | °C | °C | -- |
| （◎Д◎） |
