Template:Kyoto/Notebook/Sep 26
From 2013.igem.org
(Difference between revisions)
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(→Colony PCR) |
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|GGA-6 (1)||3359 | |GGA-6 (1)||3359 | ||
|- | |- | ||
- | |GGA-16( | + | |GGA-16(2~4)||2939 |
|} | |} | ||
</div> | </div> | ||
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|5min||30s||30s||3min30s||30cycles | |5min||30s||30s||3min30s||30cycles | ||
|} | |} | ||
- | [[File: | + | |
- | [[File: | + | ===Electrophoresis=== |
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !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)||--||-- | ||
+ | |} | ||
+ | [[File:xxxxx1.jpg]]<br> | ||
+ | </div> | ||
+ | |||
+ | {| class="wikitable" | ||
+ | !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]]<br> | ||
+ | </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]]<br> | ||
+ | </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]]<br> | ||
+ | </div> |
Revision as of 13:32, 26 September 2013
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>