From 2013.igem.org
Sep 11
PCR
Tatsui
| pKaiBC(200p) | 10*buffer KOD plus | dNTPs | MgSO4 | primer mix(F&R) | KOD plus | MilliQ | total
|
| 0.5 | 2.5 | 2.5 | 1.5 | 1.5 | 0.5 | 16 | | 25
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 98°C | 60°C | 68°C | 30
|
| 2min | 10sec | 30sec | 30sec |
|
| pKaiBC(400p) | 10*buffer KOD plus | dNTPs | MgSO4 | primer mix(F&R) | KOD plus | MilliQ | total
|
| 1 | 2.5 | 2.5 | 1.5 | 1.5 | 0.5 | 15.5 | | 25
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 98°C | 60°C | 68°C | 30
|
| 2min | 10sec | 30sec | 30sec |
|
| pKaiBC(800p) | 10*buffer KOD plus | dNTPs | MgSO4 | primer mix(F&R) | KOD plus | MilliQ | total
|
| 2 | 2.5 | 2.5 | 1.5 | 1.5 | 0.5 | 14.5 | | 25
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 98°C | 60°C | 68°C | 30
|
| 2min | 10sec | 30sec | 30sec |
|
| pKaiBC(1ng) | 10*buffer KOD plus | dNTPs | MgSO4 | primer mix(F&R) | KOD plus | MilliQ | total
|
| 2.5 | 2.5 | 2.5 | 1.5 | 1.5 | 0.5 | 14 | | 25
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 98°C | 60°C | 68°C | 30
|
| 2min | 10sec | 30sec | 30sec |
|
colony PCR
Tatsui
| Sample | base pair
|
| 9/9Pcon-lacZα(p3B4k5) | 712bp
|
|9/9Pcon-spinach-DT(p3B4k5)||605bp
|}
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 42s |
|
| Sample | base pair
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 42s |
|
| Sample | base pair
|
| Pbad\araC-RBS-luxI-DT |
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 5min | 30s | 30s | 2min |
|
| Sample | base pair
|
| 9/9Pcon+RBS-luxI-DT | 1079bp
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 50sec | 30sec | 30sec | 1min |
|
| Sample | base pair
|
| 9/10Pcon+sptamer12_1R-DT | 584bp
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 50sec | 30sec | 30sec | 1min |
|
| Sample | base pair
|
| 9/10DT+Pcon-pT181 attenuater 1 | 781bp
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 50sec | 30sec | 30sec | 1min |
|
| Sample | base pair
|
| 9/10Fusion1 attenuater(p3B1C3)-1 | 638bp
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94°C | 94°C | 55°C | 68°C | --
|
| 50sec | 30sec | 30sec | 1min |
|
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