The Jacks Lab
protocols
H-ras LSL G12D PCR
Reaction Mix:
| Tail DNA | 2.0ul |
| 10x buffer | 2.0ul |
| F2 | 0.5ul |
| R1 | 1.0ul |
| SD5 | 0.5ul |
| milliQ | 13ul |
| House Taq | 1.0ul |
PCR Primers:
| F2 | 5' cca gcc atc cct cgc gtt cc 3' |
| R1 | 5’ gca ggg ctg aca gct gag agc 3’ |
| SD5 | 5’ agc tag cca cca tgg ctt gag taa gtc tgc a 3’ |
PCR Conditions:
| 1. | 94C | 2:00 |
| 2. | 94C | 0:30 |
| 3. | 60C | 0:30 |
| 4. | 72C | 0:40 |
| 5. | Goto 2 x34 cycles | |
| 6. | 72C | 5:00 |
| 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 391 bp |
LSL |