The
Jacks Lab
protocols
Kras G12D Conditional
PCR
(multiplex & single allele)
Use Promega Green Taq Master Mix
Reaction Mix:
| DNA 100ng | 1.0ul |
| Master Mix (2x) | 5.0ul |
| Primer Mix (5uM each) | 1ul |
| milliQ | fill to 10 ul |
PCR Primers:
| 1 | 5' gtc ttt ccc cag cac agt gc 3' |
| 2 | 5' ctc ttg cct acg cca cca gct c 3’ |
| 3 | 5’ agc tag cca cca tgg ctt gag taa gtc tgc a 3’ |
PCR Conditions:
| 1. | 95C | 2:00 |
| 2. | 95C | 0:30 |
| 3. | 61C | 0:30 |
| 4. | 72C | 0:45 |
| 5. | Goto 2 x34 cycles | |
| 6. | 72C | 10:00 |
| 7. | 4C | 0:00 |
| END | ||
Expected Bands: (run gel a long way to discern between the 3)
|
Wt= ~622bp
|
LSL-cassette= 500bp
|
1 lox (recombined) = ~650bp |
What a gel would look like:
Location of Primers:
