Reaction Mix:
| Tail DNA | 2.0ul |
| 10x buffer | 2.0ul |
| primer 3′ lox | 1.0ul (20uM) |
| primer 5′ lox | 1.0ul (20uM) |
| milliQ | 13ul |
| House Taq | 1.0ul |
PCR Primers:
| 3′ lox primer: | 5′ CCT TgA CCA TAg CCC AgC AC 3′ |
| 5′ lox primer: | 5’ CTC TAg ATC CTC TCA TTC TTC 3’ |
PCR Conditions:
| 1. | 94C | 3:00 |
| 2. | 94C | 0:30 |
| 3. | 55C | 1:00 |
| 4. | 72C | 0:45 |
| 5. | Goto 2 x31 cycles | |
| 6. | 72C | 5:00 |
| 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:
| WILD TYPE 250 bp |
| LOXP 310 bp |