P53 conditional PCR (BERNS ALLELE)

P53FLOX RECOMBINED

Reaction Mix:

Tail DNA 1.0ul

primer A (5uM)

1.0 ul
primer B (5uM) 0.5 ul
primer D (20uM) 0.5 ul
milliQ 2.0 ul
GoTaq (Promega) 5.0 ul

PCR Primers:

A: 5' CAC AAA AAC AGG TTA AAC CCA G 3'
B: 5' AGC ACA TAG GAG GCA GAG AC 3'
D: 5' GAA GAC AGA AAA GGG GAG GG 3'

PCR Conditions:

1. 94C 2:00
2. 94C 0:30
3. 58C 0:30
4. 72C 0:50
5. Goto 2 x29 cycles
6. 72C 5:00
END

Expected Bands:

WILD TYPE
288 bp
FLOX
370 bp
Recombined
(1 lox)
612 bp

A background band sometimes appears around 400-420 bp.