The Jacks Lab
protocols
Rb1lox PCR -- to detect recombination
Reaction Mix:
| Tail DNA 100ng | 1.0 ul |
| D1 (20uM) | 0.5 ul |
| 1lox 5' (20uM) | 0.5 ul |
| 3' lox (20 uM) | 1 ul |
| milliQ | 2 ul |
| Go Taq (Promega) | 5 ul |
PCR Primers:
| D1 | 5' GCA GGA GGC AAA AAT CCA CAT AAC 3' |
| 1lox 5' | 5’ CTC TAG ATC CTC TCA TTC TTC CC 3’ |
| 3' lox | 5’ CCT TGA CCA TAG CCC AGC AC 3’ |
PCR Conditions:
| 1. | 94C | 3:00 |
| 2. | 94C | 0:30 |
| 3. | 55C | 1:00 |
| 4. | 72C | 1:30 |
| 5. | Goto 2 x30 cycles | |
| 6. | 72C | 5:00 |
| END | ||
Expected Bands:
| Wt 250 bp |
FLOX 310 bp |
| 1 lox ~ 500bp |