The Jacks Lab
protocols

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