Reaction Mix:
| Tail DNA | 2.0ul |
| 10x buffer | 2.0ul |
| +116F (20umol) | 1.0ul |
| 135 (20umol) | 1.0ul |
| Neo 19+ (20umol) | 1.0ul |
| milliQ | 12.0ul |
| Taq (1/25) | 1.0ul |
PCR Primers:
| +116F | 5′ AAG CCT TGA TTC TGA TGT GGG C 3′ |
| 135 | 5’ TGA CGA AGT CAA AGT TCC ACC G 3’ |
| Neo 19+ | 5’ GCT ATC AGG ACA TAG CGT TGG C 3’ |
PCR Conditions:
| 1. | 94C | 4:00 |
| 2. | 94C | 1:00 |
| 3. | 60C | 1:00 |
| 4. | 72C | 2:00 |
| 5. | Goto 2 x35 cycles | |
| 6. | 72C | 7:00 |
| 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= ~900 bp |
| Mut= ~759 bp |