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ROSACreERT2 PCR
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 |
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