The
Jacks Lab
protocols
N-ras LSL G12D
PCR
Reaction Mix:
| DNA 100ng | 2.0ul |
| PCR Buffer 10x | 2.0ul |
| Primer F2 | 0.5ul |
| Primer R1 | 1.0 ul |
| Primer SD5' | 0.5 ul |
| HouseTaq (1:10) | 1 ul |
| milliQ | 13 ul |
PCR Primers:
| F2 | 5' aga cgc gga gac ttg gcg agc 3' |
| R1 | 5' gct gga tcg tca agg cgc ttt tcc 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 | 7:00 |
| 7. | 15C | 4ever |
| END | ||
Expected Bands:
|
Wild type 500 bp
|
LSL-cassette 345 bp
|