The Jacks Lab
protocols
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.