Template:Kyoto/Notebook/Sep 25
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|} | |} | ||
</div> | </div> | ||
+ | |||
+ | ===Ligation=== | ||
+ | |||
+ | <!-- こっから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">じっけんしゃ</span> | ||
+ | {| class="wikitable" | ||
+ | !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | ||
+ | |- | ||
+ | |experiment||name||xx µL||name||xx µL||xx µL | ||
+ | |- | ||
+ | |NC||name||xx µL||name||xx µL||xx µL | ||
+ | |} | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | |||
+ | ===PCR=== | ||
+ | <!-- ここから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !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 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | | °C|| °C|| °C|| °C||-- | ||
+ | |- | ||
+ | |(◎Д◎) || || || || | ||
+ | |} | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | |||
+ | ===Transformation=== | ||
+ | <!-- こっから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">じっけんしゃ</span> | ||
+ | {| class="wikitable" | ||
+ | !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||- | ||
+ | |- | ||
+ | |いでんし||なんちゃらµL||ごにょごにょµL||ほにゃららµL||こーせーぶっしつ | ||
+ | |- | ||
+ | |いかどうよう|| || || || | ||
+ | |- | ||
+ | | || || || || | ||
+ | |} | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | |||
+ | ===Electrophoresis=== | ||
+ | |||
+ | <!-- こっから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">じっけんしゃ</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||Sample||Enzyme1||Enzyme2 | ||
+ | |- | ||
+ | |(°Д°)|| || || | ||
+ | |} | ||
+ | [[File:igku_xxxxxx.xxx]]<br> | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | |||
+ | ===精製?=== | ||
+ | |||
+ | ===Restriction Enzyme Digestion=== | ||
+ | <!-- こっから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">じっけんしゃ</span> | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | |} | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | |||
+ | ===液体培養=== | ||
+ | |||
+ | ===プレーティング=== | ||
+ | |||
+ | ===Colony PCR=== | ||
+ | <!-- ここから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |(`・ω・´)|| | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | | °C|| °C|| °C|| °C||-- | ||
+ | |- | ||
+ | |(◎Д◎) || || || || | ||
+ | |} | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | |||
+ | ===RNA抽出=== | ||
+ | |||
+ | ===cDNA合成=== | ||
+ | |||
+ | ===Electrophoresis=== | ||
+ | |||
+ | <!-- こっから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">じっけんしゃ</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||Sample||Enzyme1||Enzyme2 | ||
+ | |- | ||
+ | |(°Д°)|| || || | ||
+ | |} | ||
+ | [[File:igku_xxxxxx.xxx]]<br> | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
===Ligation=== | ===Ligation=== |
Revision as of 12:51, 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 |
---|---|
(・ω・) | |
(・X・) |
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
(>ω・)b | ||
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 |
---|---|
(・ω・) | |
(・X・) |
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 |
---|---|---|---|---|
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 | - |
いでんし | なんちゃらµL | ごにょごにょµL | ほにゃららµL | こーせーぶっしつ |
いかどうよう | ||||
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 |