|
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 |
|
