"
Template:Kyoto/Notebook/Sep 26
From 2013.igem.org
Contents |
Sep 26
Electrophoresis
| Lane | Sample | Enzyme1 | Enzyme2 | |
|---|---|---|---|---|
| 1 | GGA1-1 | |||
| 2 | GGA1-2 | |||
| 3 | GGA1-3 | |||
| 4 | GGA1-4 | |||
| 5 | GGA1-5 | |||
| 6 | GGA1-6 | |||
| 7 | GGA1-7 | |||
| 8 | GGA1-8 | |||
| 9 | 1kbp ladder | |||
| 10 | GGA2-1 | |||
| 11 | GGA2-2 | |||
| 12 | GGA2-3 | |||
| 13 | GGA2-4 | |||
| 14 | GGA2-5 | |||
| 15 | GGA2-6 | |||
| 16 | GGA2-7 | |||
| 17 | GGA2-8 |
| Lane | Sample | Enzyme1 | Enzyme2 | |
|---|---|---|---|---|
| 1 | GGA3-1 | |||
| 2 | GGA3-2 | |||
| 3 | GGA3-3 | |||
| 4 | 1kbp ladder | |||
| 5 | GGA3-4 | |||
| 6 | GGA3-5 | |||
| 7 | GGA3-6 | |||
| 8 | GGA3-7 |
File:Xxxxx.png
</div>
| Lane | Sample | Enzyme1 | Enzyme2 | |
|---|---|---|---|---|
| 1 | GGA3-8 | |||
| 2 | GGA4-1 | |||
| 3 | GGA4-2 | |||
| 4 | GGA4-3 | |||
| 5 | 1kbp ladder | |||
| 6 | GGA4-4 | |||
| 7 | GGA4-5 | |||
| 8 | GGA4-6 |
File:Xxxxx.png
</div>
| Lane | Sample | Enzyme1 | Enzyme2 | |
|---|---|---|---|---|
| 1 | GGA4-7 | |||
| 2 | GGA4-8 | |||
| 3 | 1kbp ladder | |||
| 4 | GGA7-1 | |||
| 5 | GGA16-1 | |||
| 6 | NC(RBS-GFP-DT) |
File:Xxxxx.png
</div>
Ligasion(Golden Gate Assenbly)
| state | Vector | Inserter1 | Inserter2 | Inserter3 | Inserter4 | Inserter5 | Ligation High ver.2 | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| experiment | RBS-GFP-DT | 1.0 µL | Pcon-pT181 antisense(E-2A) | 0.6µL | DT(2-1) | 0.5µL | Pcon-pT181 attenuator(1-SA) | 2.1 µL | -- | -- | -- | -- | 2.1 µL |
| experiment | pSB1C3(EcoRI&SpeI) | 1.8 µL | Pcon-pT181 antisense(E-2A) | 0.9 µL | DT(2-1) | 0.5 µL | Pcon-GFP-DT(1-SA) | 1.5 µL | -- | -- | -- | -- | 2.5 µL |
| experiment | RBS-GFP-DT | 1.0 µL | Ptet(E-0) | 0.7µL | Pcon pT181 attenuator(0-1A) | 0.3µL | tet aptamer(1-3) | 0.3 µL | Pcon-tetR-DT(3-2A) | 1.1 µL | Ptet(2-5) | 0.3 µL | 2.2 µL |
| experiment | RBS-GFP-DT | 1.0 µL | Ptet(E-0) | 0.7 µL | Pcon-pT181 attenuator-DT(0-1A) | 1.0 µL | Pcon-tetR-DT(1-2A) | 1.0 µL | Ptet(2-5) | 0.3µL | -- | -- | 2.0 µL |
| experiment | RBS-GFP-DT | 1.0 µL | Pcon-pT181 attenuator-tetR-DT(E-1A) | 1.0 µL | Pcon antisense-spinach-DT(1-2A) | 0.7µL | Pcon-pT 181 attenuator(2-SA) | 2.1 µL | -- | -- | -- | -- | 2.4 µL |
| experiment | RBS-GFP-DT | 1.0 µL | Pcon antisense(E-1A) | 0.5 µL | spinach-DT(1-2) | 1.0 µL | Pcon-GFP-DT(1-2) | 0.8 µL | -- | -- | -- | -- | 2.15 µL |
| experiment | pSB1C3(EcoRI&SpeI) | 1.8 µL | Pcon-pT181 antisense(E-1A) | 0.8µL | spinach-DT(1-2) | 1.0µL | Pcon-GFP-DT(2-SA) | 0.8 µL | -- | -- | -- | -- | 2.1 µL |
| experiment | pSB1C3(EcoRI&SpeI) | 1.8 µL | Pcon-tet aptamer-DT(E-1A) | 3.0 µL | Ptet-pT181 antisense(1-3) | 1.5 µL | spinach-DT(3-2) | 1.5 µL | Pcon-GFP-DT(2-5A) | 0.8 µL | -- | -- | 4.3 µL |
Liquid Culture
| Sample | medium |
|---|---|
| Pcon-GFP-DT | Amp |
| RBS-GFP-DT | CP |
| Pcon-pT181 attenuator-DT | Amp |
| Pcon-tet aptamer-DT | Amp |
| Pcon-spinach-DT | Amp |
| Pcon-pT181 antisense-spinach-DT | Amp |
| spinach-DT | CP |
BSAI Digestion
| DNA | MilliQ | BSAI-HF | 10*cut smart | total | |
|---|---|---|---|---|---|
| Pcon | 7 µL | 10.7 µL | 0.3 µL | 2 µL | 20 µL |
| RBS-luxR-DT | 10 µL | 7 µL | 0.3 µL | 2 µL | 20 µL |
Colony PCR
| Sample | base pair |
|---|---|
| GGA-1 (9~16) | 3177 |
| GGA-2 (9~16) | 3189 |
| GGA-3 (9~21) | 2989 |
| GGA-4 (9~16) | 2939 |
| GGA-6 (1) | 3359 |
| GGA-16(2~4) | 2939 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 3min30s | 30cycles |
Electrophoresis
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | GGA-1 (9) | -- | -- |
| 2 | GGA-1 (10) | -- | -- |
| 3 | GGA-1 (11) | -- | -- |
| 4 | GGA-1 (12) | -- | -- |
| 5 | GGA-1 (13) | -- | -- |
| 6 | GGA-1 (14) | -- | -- |
| 7 | GGA-1 (15) | -- | -- |
| 8 | GGA-1 (16) | -- | -- |
| 9 | 1kbp ladder | -- | -- |
| 10 | GGA-2 (9) | -- | -- |
| 11 | GGA-2 (10) | -- | -- |
| 12 | GGA-2 (11) | -- | -- |
| 13 | GGA-2 (12) | -- | -- |
| 14 | GGA-2 (13) | -- | -- |
| 15 | GGA-2 (14) | -- | -- |
| 16 | GGA-2 (15) | -- | -- |
| 17 | GGA-2 (16) | -- | -- |
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | GGA-4 (9) | -- | -- |
| 2 | GGA-4 (10) | -- | -- |
| 3 | GGA-4 (11) | -- | -- |
| 4 | GGA-4 (12) | -- | -- |
| 5 | GGA-4 (13) | -- | -- |
| 6 | GGA-4 (14) | -- | -- |
| 7 | GGA-4 (15) | -- | -- |
| 8 | GGA-4 (16) | -- | -- |
| 9 | 1kbp ladder | -- | -- |
| 10 | GGA-6 (1) | -- | -- |
| 11 | GGA-16 (2) | -- | -- |
| 12 | GGA-16 (3) | -- | -- |
| 13 | GGA-16 (4) | -- | -- |
File:Xxxxx1.jpg
</div>
|1||GGA-3 (9)||--||--
|-
|2||GGA-3 (10)||--||--
|-
|3||GGA-3 (11)||--||--
|-
|4||GGA-3 (12)||--||--
|-
|5||1kbp ladder||--||--
|-
|6||GGA-3 (13)||--||--
|-
|7||GGA-3 (15)||--||--
|-
|8||GGA-3 (14)||--||--
|}
File:Xxxxx1.jpg
</div>
|1||GGA-3 (17)||--||--
|-
|2||GGA-3 (18)||--||--
|-
|3||GGA-3 (19)||--||--
|-
|4||1kbp ladder||--||--
|-
|5||GGA-3 (20)||--||--
|-
|6||GGA-3 (21)||--||--
|-
|7||GGA-3 (16)||--||--
|}
File:Xxxxx1.jpg
</div>
