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:
