Reaction Mix:
| Tail DNA | 2.0 ul |
| 10x PCR buffer with NTPs | 2.0 ul |
| Flox 1 (20uM) | 1.0 ul |
| Flox 2 (20uM) | 1.0 ul |
| milliQ | 13 ul |
| House Taq (1:10) | 1.0 ul |
PCR Primers:
| Flox 1 | 5′ GAT GGG TCT GTA GTC TGG G 3′ |
| Flox 2 | 5’ GGC TCA GCG TTT TCC TAA TG 3’ |
PCR Conditions:
| 1. | 94C | 3:00 |
| 2. | 94C | 0:30 |
| 3. | 56C | 1:00 |
| 4. | 72C | 0:30 |
| 5. | Goto 2 x29 cycles | |
| 6. | 72C | 10:00 |
| 7. | 4C | 0:00 |
| END | ||
10X PCR Buffer:
| 500 mM KCl |
| 100mM Tris pH 8.3 |
| 15 mM MgCl2 |
| 1 mg/ml BSA |
| 2 mM dNTPs |
| dH20 |
(store at -20C)
Expected Bands:
| Wt = 340 bp |
| Flox = 500 bp |