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 |