NYMU Taipei/Lab Notes/2007 9 30
From 2007.igem.org
PCR for TAT_A-INS_A TAT_B-INS_B
| size | Primer | Template | ||
|---|---|---|---|---|
| 1 | TAT_A-INS_A | 209 | TAT_F+INS_A_R | TAT_A,INS_A PCR product@9/29 |
| 2 | TAT_B-INS_B | 233 | TAT_F+INS_B_R | TAT_B,INS_B PCR product@9/29 |
| Template1 | 1uL | ||
| Template2 | 1uL | ||
| 10X buffer | 5uL | ||
| 2.5mM dNTP | 2uL | ||
| Primer_F | 1.5uL | ||
| Primer_R | 1.5uL | ||
| Pfu polymerase | 0.5uL | ||
| ddH2O | 37.5uL | ||
| Total | 50uL |
| Step | Temp (°C) | Time (mins) | Cycle |
|---|---|---|---|
| Initial Denaturation | 95 | 10 | |
| Denatuation Annealing Extension | 95 55 72 | 1 1 1 | 35 |
| Final Extension | 72 | 10 | |
| Soak | 4 |
2%Gel check for TAT_A-INS_A TAT_B-INS_B
- Well 1 TAT_A-INS_A 10uL
- Well 2 TAT_B-INS_B 10uL
- Well 3 100bp ladder 5uL
