The Jacks Lab

 

protocols

Rb conditional PCR

Reaction Mix:

Tail DNA 2.0ul
10x buffer 2.0ul
primer 3' lox 1.0ul (20uM)
primer 5' lox 1.0ul (20uM)
milliQ 13ul
House Taq 1.0ul

PCR Primers:

3' lox primer: 5' CCT TgA CCA TAg CCC AgC AC 3'
5' lox primer: 5’ CTC TAg ATC CTC TCA TTC TTC 3’

PCR Conditions:

1. 94C 3:00
2. 94C 0:30
3. 55C 1:00
4. 72C 0:45
5. Goto 2 x31 cycles
6. 72C 5:00
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:

WILD TYPE

250 bp

LOXP

310 bp