Reaction Mix (store at -20C):
| 100 pmol stock NF2WT | 20 ul |
| 100 pmol stock NF2X2 | 20 ul |
| 100 pmol stock neo18 | 20 ul |
| 10 X buffer (see below) | 500 ul |
| milliQ H2O | 4430 ul |
For each reaction:
| Tail DNA | 2 ul |
| RX Mix | 17 ul |
| Taq (1/25) | 1.0ul |
PCR Primers:
| Nf2WT | 5' GTC TGG GAA GTC TGT GGA GG 3' |
| Nf2X2 | 5’ GGG GCT TCG GGA AAC CTG G 3’ |
| Neo18 | 5’ CTA TCA GGA CAT AGC GTT GG 3’ |
PCR Conditions:
| 1. | 94C | 1:00 |
| 2. | 94C | 0:30 |
| 3. | 61C | 1:45 |
| 4. | 72C | 1:45 |
| 5. | Go to 2 x29 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:
| WT = 306 bp |
| Mut = 574 bp |