Reactivatable allele as described in Ventura et al., Nature 2007
Reaction Mix:
| Tail DNA | 2.0 ul |
| MgCl2 | 0.75 ul |
| 10x PCR buffer | 2.5 ul |
| dNTPs | 0.5 ul |
| LSL Universal (100uM) | 0.05 ul |
| LSL SD5 (100uM) | 0.05 ul |
| LSL ER (100uM) | 0.05 ul |
| milliQ | 18.9 ul |
| Taq | 0.2 ul |
PCR Primers:
| LSL Universal | 5′ CTT GGA GAC ATA GCC ACA CTG 3′ |
| LSL SD5 | 5’ AGC TAG CCA CCA TGG CTT GAG TAA GT 3’ |
| LSL ER418021 | 5’ CAA CTG TTC TAC CTC AAG AGC C 3’ |
PCR Conditions:
| 1. | 94C | 3:00 |
| 2. | 94C | 0:30 |
| 3. | 60C | 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:
