|
APC flox PCR
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 |
|
