From 2013.igem.org
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Revision as of 17:54, 23 September 2013
Sep 9
PCR
Nakagawa
| KaiA(plasmid) | 10x buffer | dNTPs | MgSO4 | Primer(661-RBS-KaiA-fwd) | Primer(kaiA-662-rev-ver2) | Kod-plus ver2 | MilliQ | total
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| 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
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| 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
No name
| DNA | concentration[µg/mL] | 260/280 | 260/230
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| 9/8 pSB1C3 | 83.2 | 1.45 | 0.76
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| 9/8 RBS-lysis2-DT | 72.8 | 1.67 | 0.97
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| 9/8 spinach-DT | 152.6 | 1.88 | 1.97
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| Nuclease-Free water | -0.4 | -14.30 | 0.25
|
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