Template:Kyoto/Notebook/Sep 11
From 2013.igem.org
Sep 11
PCR
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
Sample | base pair |
---|---|
Pcon-lacZα(p3B4k5) | 712bp |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 42s |
Sample | base pair |
---|---|
Pcon-spinach-DT(p3B4k5) | 605bp |
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 | 605bp |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 2min |