Template:Kyoto/Notebook/Sep 25
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!Lane||Sample||Enzyme1||Enzyme2 | !Lane||Sample||Enzyme1||Enzyme2 | ||
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- | | | + | |1||100bp ladder|| || |
+ | |- | ||
+ | |2||Pcon-RBS-GFP-DT(RNA)|| || | ||
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[[File:igku_xxxxxx.xxx]]<br> | [[File:igku_xxxxxx.xxx]]<br> |
Revision as of 14:05, 26 September 2013
Sep 25
Colony PCR
Sample | base pair |
---|---|
9/23 Pcon-PT181 attenuator-aptamer12-1R-DT-(5~12) | 859 |
9/23 Pcon-PT181 attenuator-RBS-GFP-DT-(5~8) | 1527 |
9/23 Pcon-PT181 attenuator-pSB1C3-(5~8) | 601 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94 °C | 94 °C | 55 °C | 68 °C | -- |
2 min | 30 s | 30 s | 1min5s | 30 cycles |
Electrophoresis
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | Pcon-PT181 attenuator-aptamer12-1R-DT-5 | -- | -- |
2 | Pcon-PT181 attenuator-aptamer12-1R-DT-6 | -- | -- |
3 | Pcon-PT181 attenuator-aptamer12-1R-DT-7 | -- | -- |
4 | Pcon-PT181 attenuator-aptamer12-1R-DT-8 | -- | -- |
5 | Pcon-PT181 attenuator-aptamer12-1R-DT-9 | -- | -- |
6 | Pcon-PT181 attenuator-aptamer12-1R-DT-10 | -- | -- |
7 | Pcon-PT181 attenuator-aptamer12-1R-DT-11 | -- | -- |
8 | Pcon-PT181 attenuator-aptamer12-1R-DT-12 | -- | -- |
9 | 1kbp ladder | -- | -- |
10 | Pcon-PT181 attenuator-RBS-GFP-DT-5 | -- | -- |
11 | Pcon-PT181 attenuator-RBS-GFP-DT-6 | -- | -- |
12 | Pcon-PT181 attenuator-RBS-GFP-DT-7 | -- | -- |
13 | Pcon-PT181 attenuator-RBS-GFP-DT-8 | -- | -- |
14 | Pcon-PT181 attenuator-pSB1C3-5 | -- | -- |
15 | Pcon-PT181 attenuator-pSB1C3-6 | -- | -- |
16 | Pcon-PT181 attenuator-pSB1C3-7 | -- | -- |
17 | Pcon-PT181 attenuator-pSB1C3-8 | -- | -- |
Transformation
Name | Sample | Competent Cells | Total | Plate |
---|---|---|---|---|
Plac+tet aptamer12+Pcon tetR-DT+Ptet+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-pT181attenuator-DT+Pcon-tetR-DT+Ptet+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-tet aptamer12-DT+Pcon-tetR-DT+Ptet+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-Spinach-DT+Pcon-tetR-DT+Ptet+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-pT181antisense+DT+Pcon-tetR-DT+Ptet+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-pT181attenuator+tet aptamer12-DT+Pcon-tetR-DT+Ptet+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon tetR-DT+Ptet+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon tetR-DT+Pcon-GFP-DT+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-tet aptamer12-DT+Pcon-pT181attenuator+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-pT181attenuator-DT+Pcon-pT181attenuator+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-tet aptamer12-DT+Ptet-pT181antisense+Spinach-DT+Pcon-RBS-GFP-DT+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-tet aptamer12-DT+Pcon-pT181antisense+Spinach-DT+Pcon-pT181attenuator+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Pcon-tet aptamer12-DT+Ptet-pT181antisense+Spinach-DT+Pcon-pT181attenuator+RBS-GFP-DT | 1µL | 10µL | 11µL | - |
Name | Sample | Competent Cells | Total | Plate |
---|---|---|---|---|
aptamer12-DT(EcoRI+Xbal)+Pcon-pT181attenuator(EcoRI+Spel) | 2µL | 20µL | 22µL | - |
Ptet(Spel+Pstl)+RBS-GFP-DT(Xbal+Pstl)) | 2µL | 20µL | 22µL | - |
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | 9/25 aptamer12_1R-DT(EcoRI&XbaI) | 4.5 µL | Pcon-pT181 attenuator(EcoRI&SpeI) | 7.7 µL | 6.1 µL |
experiment | 9/17 pSB1C3(EcoRI&SpeI) | 2.0 µL | 9/3 pT181 attenuator(EcoRI&SpeI) | 14 µL | 8 µL |
experiment | 9/25 Pcon-pT181 attenuator(EcoRI&XbaI) | 4.5 µL | 9/25 RBS-GFP-DT(EcoRI&SpeI) | 19.3 µL | 11.9 µL |
experiment | 9/25 Pcon-RBS-GFP-DT(EcoRI&XbaI) | 4.2 µL | Pcon-RBS-tetR-DT(EcoRI&SpeI) | 20 µL | 12.1 µL |
Digestion
Liquid Culture
Sample | medium |
---|---|
DT | - |
pT181attenuator | - |
Pcon-RBS-tetR-DT | - |
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | - |
2 | Pcon-pT181antisense | |
3 | ||
4 | ||
5 | - | |
6 | DT | |
7 | ||
8 | ||
9 | - | |
10 | Spinach-DT | |
11 | ||
12 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(´ω`) | |||
(ΦωΦ^) |
Lane | DNA | Enzyme |
---|---|---|
1 | ||
2 | ||
3 | ||
4 | ||
5 | ||
6 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(´ω`) | |||
(ΦωΦ^) |
Lane | DNA | Enzyme |
---|---|---|
1 | ||
2 | ||
3 | ||
4 | ||
5 | ||
6 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(´ω`) | |||
(ΦωΦ^) |
Lane | DNA | Enzyme |
---|---|---|
1 | ||
2 | ||
3 | ||
4 | ||
5 | ||
6 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(´ω`) | |||
(ΦωΦ^) |
Lane | DNA | Enzyme |
---|---|---|
1 | ||
2 | ||
3 | ||
4 | ||
5 | ||
6 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(´ω`) | |||
(ΦωΦ^) |
Lane | DNA | Enzyme |
---|---|---|
1 | ||
2 | ||
3 | ||
4 | ||
5 | ||
6 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(´ω`) | |||
(ΦωΦ^) |
Restriction Enzyme Digestion
DNA | restricion enzyme | restriction enzyme | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | (°°) µL | µL | µL | µL | µL | µL | µL |
NC | µL | µL | µL | µL | µL | µL | µL |
Liquid Culture
Sample | medium |
---|---|
Pcon-pT181anntisense-spinach-DT(4k5) | - |
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | name | xx µL | name | xx µL | xx µL |
NC | name | xx µL | name | xx µL | xx µL |
PCR
genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total |
---|---|---|---|---|---|---|---|---|
0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 25 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
°C | °C | °C | °C | -- |
(◎Д◎) |
Transformation
Name | Sample | Competent Cells | Total | Plate |
---|---|---|---|---|
aptamer12-1R-DT | µL | µL | µL | CP |
pSB1C3(EcoRI+Spel)+pT181attenuator(EcoRI+Spal) | µL | µL | µL | CP |
Pcon-pT181attenuator(EcoRI+Xbal)+RBS-GFP-DT(EcoRI+Spel) | µL | µL | µL | Amp |
Pcon-RBS-GFP-DT(EcoRI+Xbal)+Pcon-RBS-tetR-DT(EcoRI+Spel) | µL | µL | µL | Amp |
pSB1C3(EcoRI+Spel)+Pcon-pT181attenuator-aptamer12-1R-DT(EcoRI+Spel) | µL | µL | µL | CP |
Electrophoresis
精製?
Restriction Enzyme Digestion
DNA | restricion enzyme | restriction enzyme | Buffer | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | (°°) µL | µL | µL | µL | µL | µL | µL |
NC | µL | µL | µL | µL | µL | µL | µL |
液体培養
プレーティング
Colony PCR
Sample | base pair |
---|---|
(`・ω・´) |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
°C | °C | °C | °C | -- |
(◎Д◎) |
RNA抽出
cDNA合成
Electrophoresis
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | name | xx µL | name | xx µL | xx µL |
NC | name | xx µL | name | xx µL | xx µL |