Template:Kyoto/Notebook/Sep 17
From 2013.igem.org
(Difference between revisions)
(→Gel Extraction) |
(→PCR) |
||
Line 163: | Line 163: | ||
<!-- ここから --> | <!-- ここから --> | ||
<div class="experiment"> | <div class="experiment"> | ||
- | <span class="author"> | + | <span class="author">Kinose</span> |
{| class="wikitable" | {| class="wikitable" | ||
- | ! | + | !pC KaiABC(100pg)||Prime STAR(2x)||Primer(KaiA-Bsai MuT-Fwd)||Primer(KaiA-Bsai MuT-Rev)||MilliQ||total |
|- | |- | ||
- | | | + | |1.0||25.0||1.0||1.0||22.0||50.0 |
|} | |} | ||
{| class="wikitable" | {| class="wikitable" | ||
!PreDenature||Denature||Annealing||Extension||cycle | !PreDenature||Denature||Annealing||Extension||cycle | ||
|- | |- | ||
- | | | + | |96°C||98°C||65°C||72°C||-- |
|- | |- | ||
- | | | + | |5m||10s||15s||40s||30cycles |
+ | |} | ||
+ | </div> | ||
+ | <div class="experiment"> | ||
+ | {| class="wikitable" | ||
+ | !pC KaiABC(150pg)||Prime STAR(2x)||Primer(KaiA-Bsai MuT-Fwd)||Primer(KaiA-Bsai MuT-Rev)||MilliQ||total | ||
+ | |- | ||
+ | |1.5||25.0||1.0||1.0||21.5||50.0 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |96°C||98°C||65°C||72°C||-- | ||
+ | |- | ||
+ | |5m||10s||15s||40s||30cycles | ||
+ | |} | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | </div> | ||
+ | <div class="experiment"> | ||
+ | {| class="wikitable" | ||
+ | !pC KaiABC(200pg)||Prime STAR(2x)||Primer(KaiA-Bsai MuT-Fwd)||Primer(KaiA-Bsai MuT-Rev)||MilliQ||total | ||
+ | |- | ||
+ | |2||25.0||1.0||1.0||21.0||50.0 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |96°C||98°C||65°C||72°C||-- | ||
+ | |- | ||
+ | |5m||10s||15s||40s||30cycles | ||
|} | |} | ||
</div> | </div> | ||
<!-- ここまでをコピーしてね --> | <!-- ここまでをコピーしてね --> |
Revision as of 02:39, 24 September 2013
Contents |
Sep 17
Liquid Culture
Sample | medium |
---|---|
Spi-DT | LB medium(+CP) |
Pcon-Spi-DT,Pcon-pT181-anti-spi-DT | LB medium(+Amp) |
PCR
Pcon-tet ap-DT(VF) | BigDye | 5x buffer | primer(VF) | MilliQ | total |
---|---|---|---|---|---|
1.3 | 0.5 | 1.75 | 0.5 | 0.45 | 10.5 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
96°C | 96°C | 50°C | 60°C | -- |
2m | 10s | 5s | 4m | 30cycles |
Pcon-tet ap-DT(VR) | BigDye | 5x buffer | primer(VR) | MilliQ | total |
---|---|---|---|---|---|
1.3 | 0.5 | 1.75 | 0.5 | 0.45 | 10.5 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
96°C | 96°C | 50°C | 60°C | -- |
2m | 10s | 5s | 4m | 30cycles |
Ligation
state | Vector | Inserter | Ligation High ver.2 | ||
---|---|---|---|---|---|
8/28 | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbpladder | -- |
2 | Pt181 atte -(2) | E+S |
3 | ||
4 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
PT181-atte E+S | 9.2 | 1.15 | 0.12 |
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbpladder | -- |
2 | pSB1C3 | E+S |
3 | ||
4 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
pSB1C3 | 10.0 | 1.84 | 0.40 |
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbpladder | -- |
2 | pSB4K5 | X+P |
3 | ||
4 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
pSB4K5 | 7.6 | 1.81 | 0.32 |
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbpladder | -- |
2 | Pcon-RBS-tetR-DT | X+P |
3 | ||
4 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
Pcon-RBS-tetR-DT | 8.0 | 2.40 | 1.14 |
Lane | DNA | Enzyme |
---|---|---|
1 | 1kbpladder | -- |
2 | pSB1C3 | E+P |
3 | ||
4 |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
pSB1C3 | 9.1 | 2.17 | 0.35 |
PCR
pC KaiABC(100pg) | Prime STAR(2x) | Primer(KaiA-Bsai MuT-Fwd) | Primer(KaiA-Bsai MuT-Rev) | MilliQ | total |
---|---|---|---|---|---|
1.0 | 25.0 | 1.0 | 1.0 | 22.0 | 50.0 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
96°C | 98°C | 65°C | 72°C | -- |
5m | 10s | 15s | 40s | 30cycles |
pC KaiABC(150pg) | Prime STAR(2x) | Primer(KaiA-Bsai MuT-Fwd) | Primer(KaiA-Bsai MuT-Rev) | MilliQ | total |
---|---|---|---|---|---|
1.5 | 25.0 | 1.0 | 1.0 | 21.5 | 50.0 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
96°C | 98°C | 65°C | 72°C | -- |
5m | 10s | 15s | 40s | 30cycles |
</div>
pC KaiABC(200pg) | Prime STAR(2x) | Primer(KaiA-Bsai MuT-Fwd) | Primer(KaiA-Bsai MuT-Rev) | MilliQ | total |
---|---|---|---|---|---|
2 | 25.0 | 1.0 | 1.0 | 21.0 | 50.0 |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
96°C | 98°C | 65°C | 72°C | -- |
5m | 10s | 15s | 40s | 30cycles |