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Template:Kyoto/Notebook/Sep 9
From 2013.igem.org
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| + | ===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 --> | ||
Revision as of 18:30, 23 September 2013
Contents |
Sep 9
PCR
| KaiA(plasmid) | 10x buffer | dNTPs | MgSO4 | Primer(661-RBS-KaiA-fwd) | Primer(kaiA-662-rev-ver2) | Kod-plus ver2 | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 1.0 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.5 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 98°C | 58.5°C | 68°C | -- |
| 2min | 10s | 30s | 30s | 30cycles |
| KaiB(plasmid) | 10x buffer | dNTPs | MgSO4 | Primer(661-RBS-KaiA-fwd-ver2) | Primer(kaiB-663-rev-ver2) | Kod-plus ver2 | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 1.0 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.5 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 98°C | 58.5°C | 68°C | -- |
| 2min | 10s | 30s | 30s | 30cycles |
| P-KaiBC(PCR products) | 10x buffer | dNTPs | MgSO4 | Primer(prefix-P-kaiBC-fwd-ver2) | Primer(P-KaiBC-suffix-rev-verw) | Kod-plus ver2 | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 0.7 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.8 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 98°C | 58.5°C | 68°C | -- |
| 2min | 10s | 30s | 30s | 30cycles |
| DT(plasmid) | 10x buffer | dNTPs | MgSO4 | Primer(664-DT-fwd-ver2) | Primer(DT-suffix-660-rev-ver2) | Kod-plus ver2 | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 1.0 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 0.5 | 15.5 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 98°C | 58.5°C | 68°C | -- |
| 2min | 10s | 30s | 30s | 30cycles |
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| 9/8 pSB1C3 | 83.2 | 1.45 | 0.76 |
| 9/8 RBS-lysis2-DT | 72.8 | 1.67 | 0.97 |
| 9/8 spinach-DT | 152.6 | 1.88 | 1.97 |
| Nuclease-Free water | -0.4 | -14.30 | 0.25 |
Restriction Enzyme Digestion
| DNA | restricion enzyme | restriction enzyme | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | （°°） µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL |
Transformation
| Name | Sample | Competent Cells | Total | Plate |
|---|---|---|---|---|
| いでんし | なんちゃらµL | ごにょごにょµL | ほにゃららµL | こーせーぶっしつ |
| いかどうよう | ||||
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
| NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
Colony PCR
| Sample | base pair |
|---|---|
| 9/8 Plac-spinach-DT-(1) | 659 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5m | 30s | 30s | 40s | 30cycles |
| Sample | base pair |
|---|---|
| 9/8 Plac-spinach-DT-(2) | 659 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5m | 30s | 30s | 40s | 30cycles |
| Sample | base pair |
|---|---|
| 9/8 Pcon-pt181atte-DT-(1) | 584 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5m | 30s | 30s | 40s | 30cycles |
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
| NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
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◎） |
