Template:Kyoto/Notebook/Sep 6
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==Sep 6== | ==Sep 6== | ||
===Electrophoresis=== | ===Electrophoresis=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Hirano</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||Sample | ||
+ | |- | ||
+ | |1||100bp ladder | ||
+ | |- | ||
+ | |2||9/4 RBS-lysis3-DT -3 | ||
+ | |- | ||
+ | |3||9/4 RBS-lysis3-DT -4 | ||
+ | |- | ||
+ | |4||9/4 RBS-lysis3-DT -5 | ||
+ | |- | ||
+ | |5||9/4 RBS-lysis1-DT -3 | ||
+ | |- | ||
+ | |6||9/4 Ptrc-KaiC -1 | ||
+ | |- | ||
+ | |7||9/4 pSB4K5 -1 | ||
+ | |- | ||
+ | |8||100bp ladder | ||
+ | |} | ||
+ | [[File:igku_xxxxxx.xxx]]<br> | ||
+ | </div> | ||
+ | |||
===Colony PCR=== | ===Colony PCR=== | ||
<div class="experiment"> | <div class="experiment"> |
Revision as of 15:25, 24 September 2013
Contents |
Sep 6
Electrophoresis
Lane | Sample |
---|---|
1 | 100bp ladder |
2 | 9/4 RBS-lysis3-DT -3 |
3 | 9/4 RBS-lysis3-DT -4 |
4 | 9/4 RBS-lysis3-DT -5 |
5 | 9/4 RBS-lysis1-DT -3 |
6 | 9/4 Ptrc-KaiC -1 |
7 | 9/4 pSB4K5 -1 |
8 | 100bp ladder |
Colony PCR
Sample | base pair |
---|---|
Pconst-spinach-DT-(1~4) | 563 |
Plac-spinach-DT-(1~4) | 659 |
Pcon-PT181 antisense-(1~4) | 468 |
Ptet-PT181 antisense-(1~4) | 604 |
Pconst-PT181 attenuater-(1~4) | 568 |
Pconst+RBS-tetR-DT-(1~4) | 1121 |
Ptet+RBS-lacZα-DT-(1~4) | 1162 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 40s | 30cycles |
Liquid Culture
Sample | medium |
---|---|
9/4 RBS-Lysis3+DT3 | Plusgrow medium(+Cp) |
9/4 RBS-Lysis1+DT3 | Plusgrow medium(+Cp) |
9/4 PSB4K5 | Plusgrow medium(+K) |
Colony PCR
Sample | base pair |
---|---|
Pconst-spinach-DT-(1) | - |
Pconst-spinach-DT-(2) | - |
Pconst-spinach-DT-(3) | - |
Pconst-spinach-DT-(4) | - |
Plac-spinach-DT-(1) | - |
Plac-spinach-DT-(2) | - |
Plac-spinach-DT-(3) | - |
Plac-spinach-DT-(4) | - |
Pcon-PT181 antisense-(1) | - |
Pcon-PT181 antisense-(2) | - |
Pcon-PT181 antisense-(3) | - |
Pcon-PT181 antisense-(4) | - |
Sample | base pair |
---|---|
Ptet-PT181 antisense-(1) | - |
Ptet-PT181 antisense-(2) | - |
Ptet-PT181 antisense-(3) | - |
Ptet-PT181 antisense-(4) | - |
Plac-PT181 attenuater-(1) | - |
Plac-PT181 attenuater-(2) | - |
Plac-PT181 attenuater-(3) | - |
Plac-PT181 attenuater-(4) | - |
Miniprep
DNA | concentration [µg/mL] | 260/280 | 260/230 |
---|---|---|---|
PT181 antisense-(1) | 238.7 | 1.68 | 1.44 |
PT181 antisense-(2) | 248.8 | 1.83 | 1.77 |
PT181 antisense-(3) | 223.2 | 1.78 | 1.61 |
PT181 antisense-(4) | 220.8 | 1.89 | 1.92 |
tetR aptamer-(1) | 252.5 | 1.89 | 1.90 |
tetR aptamer-(2) | 211.2 | 1.90 | 1.99 |
DT181 attenuater | 102.2 | 1.98 | 2.17 |
121R aptamer-DT | 251.7 | 1.92 | 2.01 |
PSB4K5 | 212.9 | 1.80 | 1.90 |
RBS-lacZα-DT | 185.7 | 1.78 | 1.79 |
RBS-lysis2-DT-(1) | 84.8 | 1.85 | 2.24 |
Restriction Enzyme Digestion
9/6 RBS-lysis2-DT (78.3 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|---|---|
2 cuts | 22µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 0µL | 30µL |
NC | 1.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.7µL | 10µL |
- at 37°C, for 1h
aptamer-DT (251.7 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|---|---|
2 cuts | 0.4µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 14.1µL | 30µL |
NC | 0.4µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
- at 37°C, for 1h
Pλ-luxI (86.4 µg/mL) | EcoRI | SpeI | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|---|---|
2 cuts | 23.1µL | 1µL | 1µL | 0µL | 0µL | 3µL | 3µL | 1.9µL | 30µL |
NC | 1.2µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 6.8µL | 10µL |
- at 37°C, for 1h
Colony PCR
Sample | base pair |
---|---|
Pconst+RBS-tetR-DT-(1) | 1121 |
Pconst+RBS-tetR-DT-(2) | 1121 |
Pconst+RBS-tetR-DT-(3) | 1121 |
Pconst+RBS-tetR-DT-(4) | 1121 |
Ptet+RBS-lacZα-DT-(1) | 1162 |
Ptet+RBS-lacZα-DT-(2) | 1162 |
Ptet+RBS-lacZα-DT-(3) | 1162 |
Ptet+RBS-lacZα-DT-(4) | 1162 |
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | -- |
2 | RBS-lysis2-DT | XbaI & PstI |
3 | ||
4 | -- | -- |
5 | aptamer 12-1R-DT | XbaI & PstI |
6 |
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | -- |
2 | Pλ-lux3 | EcoRI & SpeI |
3 | ||
4 | ||
5 | ||
6 |
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
RBS-lysis2-DT (XbaI&PstI) | 7.1 | 1.88 | 0.07 |
aptamer 12-1R-DT (XbaI&PstI) | 15.3 | 1.93 | 0.70 |
Pλ-lux3 (EcoRI&SpeI) | 4.5 | 1.79 | 0.31 |
Liquid Culture
Sample | medium |
---|---|
9/5 Pconst-spinach-DT-(1) | Plusgrow medium(+Amp&CP) |
9/5 Ptet-PT181 antisense-(1) | Plusgrow medium(+CP) |
9/5 Pconst+RBS-tetR-DT-(1) | Plusgrow medium(+Amp&CP) |
9/5 Ptet+RBS-lacZα-DT-(1) | Plusgrow medium(+CP) |
9/5 Pcon-PT181 attenuater-(1) | Plusgrow medium(+CP) |
9/4 pT181 antisense-(1) | Plusgrow medium(+CP) |
9/4 aptamer 12-1R-(1) | Plusgrow medium(+CP) |
Colony PCR
Sample | base pair |
---|---|
9/5 Plac-spinach-DT-(5~12) | 563 |
9/5 Pcon-antisense-(5~12) | 468 |
9/5 Plac-attenuater-(5~12) | 664 |
9/5 PT181 antisense+Plac-(1) | - |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 42s | 30cycles |