p53 R172H and R270H recombined point mutant alleles (germline) and Ser23Ala point mutant allele
Reaction Mix:
| Tail DNA | 1.0 µL |
| 10x buffer | 2.0ul |
| DT020200.1 (40uM) | 0.5µL |
| DT011200.3 (40uM) | 0.5µL |
| dH20 | 15 µL |
| Taq (1/25) | 1.0ul |
PCR Primers:
| DT020200.1 | 5' AGC CTG CCT AGC TTC CTC AGG 3' |
| DT011200.3 | 5' CTT GGA GAC ATA GCC ACA CTG 3' |
PCR Conditions:
| 1. | 94C | 3:00 |
| 2. | 94C | 1:00 |
| 3. | 60C | 2:00 |
| 4. | 72C | 2:00 |
| 5. | Goto 2 x30 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:
| Wild Type = ~290 bp |
| 1 Lox = ~330 bp |