Template:Kyoto/Notebook/Sep 27
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Revision as of 06:42, 27 September 2013
Contents |
Sep 27
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbp ladder | -- |
2 | pSB1C3 | EcoRI+SpeI |
3 | ||
4 | ||
6 | Pcon-attenuator-aptmer-DT | EcoRI+SpeI |
7 | ||
8 | ||
10 | Pcon-attenuator-aptmer-DT | EcoRI+SpeI |
11 | ||
12 |
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
Pcon-attenuator-DT(EcoRI+SpeI) | 6.8 | 1.98 | 0.43 |
pT181-attenuator(EcoRI+SpeI) | 7.5 | 1.84 | 0.06 |
pSB1C3(EcoRI+SpeI) | 17.7 | 1.97 | 0.71 |
Gel Extraction
Electrophoresis
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | pSB4K5 | EcoRI | PstI |
2 | pSB4K5 | - | - |
3 | 1kbp ladder | - | - |
4 | Pcon-attenuator | EcoRI | SpeI |
5 | Pcon-attenuator | - | - |
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | RBS-GFP-DT | XbaI | PstI |
2 | RBS-GFP-DT | - | - |
3 | RBS-GFP-DT | EcoRI | XbaI |
4 | 1kbp ladder | - | - |
5 | Ptet | EcoRI | SpeI |
6 | Ptet | - | - |
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | -- |
3 | Ptet | E+S |
4 | Ptet | E+S |
5 | Ptet | E+S |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbp ladder | -- |
3 | Pcon-antisense-aptamer-DT | X+P |
4 | Pcon-antisense-aptamer-DT | X+P |
5 | Pcon-antisense-aptamer-DT | X+P |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbp ladder | -- |
3 | RBS-GFP-DT | E+X |
4 | RBS-GFP-DT | E+X |
5 | RBS-GFP-DT | E+X |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Lane | DNA | Enzyme |
---|---|---|
1 | 1kb ladder | -- |
2 | Pcon-attenuator | E+S |
3 | Pcon-attenuator | E+S |
4 | Pcon-attenuator | E+S |
6 | pSB4K5 | E+P |
7 | pSB4K5 | E+P |
8 | pSB4K5 | E+P |
10 | RBS-GFP-DT | X+P |
11 | RBS-GFP-DT | X+P |
12 | RBS-GFP-DT | X+P |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(´ω`) | |||
(ΦωΦ^) |
Liquid Culture
Sample | medium |
---|---|
Pcon-GFP | Amp |
RBS-GFP-DT | CP |
Pcon-attenuator | Amp |
Pcon-attenuator-aptamer-DT | Amp |
Pcon-tetR-DT | Amp |
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
experiment | 9/17 pSB1C3 (EcoRI+SpeI) 10.0ng/µL | 10µL | 9/26 pT181 attenuator(EcoRI+SpeI) 7.5ng/µL | 1.3µL | 3.5 µL |
experiment | 9/25 DT (EcoRI+XbaI) 42.8ng/µL | 2.3µL | 9/22 Pcon-pT181 antisense(EcoRI+SpeI) 46.7ng/µL | 1.5µL | 1.9 µL |
experiment | 9/27 pSB4K5(EcoRI+PstI) 12.9ng/µL | 7.8µL | 9/10 Pcon-attenuator(EcoRI+SpeI) 13.8ng/µL | 8.0µL | 3.5 µL |
experiment | 9/27 pSB4K5(EcoRI+PstI) 12.9ng/µL | 7.8µL | 9/25 RBS-GFP-DT(XbaI+PstI) 13.9ng/µL | 12µL | 3.5 µL |
experiment | 9/8 pSB4K5 (EcoRI+SpeI) 17.6ng/µL | xx µL | 8/21 Pcon-GFP-DT(EcoRI+SpeI) 11ng/µL | 12.6µL | xx µL |
Liquid Culture
Sample | medium |
---|---|
Pcon-spinach-DT | plusgrow Amp |
Pcon-tetRaptamer-DT | plusgrow Amp |
Pcon-antisense-spinach-DT | plusgrow Amp |
spinach-DT | plusgrow CP |
RNA Extraction
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
9/26 Pcon-GFP-DT | 423.0 | 1.83 | 1.46 |
9/26 Pcon-GFP-DT K | 298.7 | 1.82 | 1.50 |
Electrophoresis
Miniprep
DNA |
---|
Pcon-aptamer-DT |
Pcon-spinach-DT |
Pcon-antisense |
Pcon-tetR-DT |
Pcon-attenuator-DT |
Pcon-antisense-spinach-DT |
Transformation
Name1 | Name2 | Sample1(µL) | Sample2(µL) | Competent Cells(µL) | Total(µL) |
---|---|---|---|---|---|
Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-attenuator-DT(1A2) | 2 | 1 | 30 | 33 |
Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-antisense-spinach-DT(1A2) | 2 | 1 | 30 | 33 |
Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-tetR aptamer-DT(1A2) | 2 | 1 | 30 | 33 |
Pcon-RBS-GFP-DT(4K5) | Pcon-attenuator-DT(1A2) | 2 | 1 | 30 | 33 |
Pcon-RBS-GFP-DT(4K5) | Pcon-antisense-spinach-DT(1A2) | 2 | 1 | 30 | 33 |
Pcon-RBS-GFP-DT(4K5) | Pcon-tetR aptamer-DT(1A2) | 2 | 1 | 30 | 33 |
Pcon-attenuator+RBS-GFP-DT(4K5) | -- | 2 | -- | 20 | 22 |
Pcon-RBS-GFP-DT(4K5) | -- | 2 | -- | 20 | 22 |
attenuator(1C3) | -- | 2 | -- | 20 | 22 |