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 |