Reaction Mix:
| Tail DNA | 2.0 ul |
| 10x PCR buffer | 2.0 ul |
| R26F2 (20uM) | 0.5 ul |
| R523 (20uM) | 0.5 ul |
| CreER – R1 (20uM) | 0.5 ul |
| milliQ | 13.5 ul |
| Taq (1:10) | 1 ul |
PCR Primers:
| R26F2 | 5′ AAA GTC GCT CTG AGT TGT TAT 3′ |
| R523 | 5’ GGA GCG GGA GAA ATG GAT ATG 3’ |
| CreER – R1 | 5’ CCT GAT CCT GGC AAT TTC G 3’ |
PCR Conditions:
| 1. | 94C | 3:00 |
| 2. | 94C | 0:30 |
| 3. | 58C | 1:30 |
| 4. | 72C | 1:00 |
| 5. | Goto 2 x34 cycles | |
| 6. | 72C | 5:00 |
| 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 = 650 bp |
| CreER = ~800 bp |