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Template:Kyoto/Notebook/Sep 17
From 2013.igem.org
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|pSB1C3||9.1||2.17||0.35 | |pSB1C3||9.1||2.17||0.35 | ||
|} | |} | ||
| + | <!-- ここまでをコピーしてね --> | ||
| + | ===PCR=== | ||
| + | <!-- ここから --> | ||
| + | <div class="experiment"> | ||
| + | <span class="author">No name</span> | ||
| + | {| class="wikitable" | ||
| + | !genome DNA||KOD plus||10x buffer||dNTP||MgSO4||SasA_fwd primer||SasA_rev primer||MilliQ||total | ||
| + | |- | ||
| + | | ||0.5||2.5||2.5||1.5||0.75||0.75|| ||25 | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | !PreDenature||Denature||Annealing||Extension||cycle | ||
| + | |- | ||
| + | | °C|| °C|| °C|| °C||-- | ||
| + | |- | ||
| + | |（◎Д◎） || || || || | ||
| + | |} | ||
| + | </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 02:31, 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
| genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| °C | °C | °C | °C | -- |
| （◎Д◎） |
