NYMU Taipei/Lab Notes/2007 9 29
From 2007.igem.org
PCR for TAT_A INS_A TAT_B INS_B TAT_D OmpRBS
| size | Primer | Template | ||
|---|---|---|---|---|
| 1 | TAT_A | 140bp | TAT_F+TAT_A_R | E.coli K12 genome DNA |
| 2 | INS_A | 111bp | INS_A_F+INS_A_R | MGC clone (H_INS 1) |
| 3 | TAT_B | 137bp | TAT_F+TAT_B_R | E.coli K12 genome DNA |
| 4 | INS_B | 135bp | INS_B_F+INS_B_R | MGC clone (H_INS 1) |
| 5 | TAT_D | 131bp | TAT_F+TAT_D_R | E.coli K12 genome DNA |
| 6 | OmpRBS | 100bp | OmpRBS_F+OmpRBS_R | pOmpC |
| Template | 1uL | ||
| 10X buffer | 5uL | ||
| 2.5mM dNTP | 2uL | ||
| Primer_F | 1.5uL | ||
| Primer_R | 1.5uL | ||
| pfu polymerase | 0.5uL | ||
| ddH2O | 38.5uL | ||
| 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 |
PCR for IDE
| size | Primer | Template | |
|---|---|---|---|
| IDE | 3105bp | IDE_F+IDE_R | M_IDE |
| Template | 1uL | ||
| 10X buffer | 5uL | ||
| 2.5mM dNTP | 2uL | ||
| Primer_F | 1.5uL | ||
| Primer_R | 1.5uL | ||
| pfu polymerase | 0.5uL | ||
| ddH2O | 38.5uL | ||
| Total | 50uL |
| Step | Temp (°C) | Time (mins) | Cycle |
|---|---|---|---|
| Initial Denaturation | 95 | 10 | |
| Denatuation Annealing Extension | 95 55 72 | 1 1 4 | 35 |
| Final Extension | 72 | 10 | |
| Soak | 4 |
