Reaction Mix:
Tail DNA | 2.0ul |
10x buffer | 2.0ul |
p107-NEO (20uM) | 0.2ul |
p107-WT (20uM) | 0.2ul |
p107-COM (20uM) | 0.4ul |
milliQ | 14.4ul |
Taq (1/25) | 1.0ul |
PCR Primers:
p107-NEO | 5' ACG AGA CTA GTG AGA CGT GC 3' |
p107-WT | 5’ TGT CCT GAG CAT GAA CAG AC 3’ |
p107-COM | 5’ TCG CTG GCA GTC TGA GTC AG 3’ |
PCR Conditions:
1. | 94C | 3:00 |
2. | 94C | 0:30 |
3. | 58C | 1:00 |
4. | 72C | 0:45 |
5. | Goto 2 x34 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= ~280 bp |
Mut= ~360 bp |