The Jacks Lab
protocols
Rb PCR
Reaction Mix:
| Tail DNA | 2.0ul |
| 10x buffer | 2.0ul |
| Rb-x3 | 0.4ul |
| Rb-I3 | 0.2ul |
| Rb-PGK-3 | 0.2ul |
| milliQ | 14.4ul |
| Taq (1/25) | 1.0ul |
PCR Primers:
| Rb-x3 | 5' GCA TCT GCA TCT TTA TCG C 3' |
| Rb-I3 | 5’ CCC ATG TTC GGT CCC TAG 3’ |
| Rb-PGK-3 | 5’ GAA GAA CGA GAT CAG CAG C 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 |
| 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= ~380 bp |
| Mut= ~410 bp |