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Kras KO PCR
Reaction Mix:
| Tail DNA |
4.0ul |
| 10x PCR buffer |
2.0ul |
| k-5'-IO (20uM) |
0.2ul |
| K-3'-1a (20uM) |
0.1ul |
| K-3'-neo (20uM) |
0.25ul |
| milliQ |
12.45ul |
| Taq (1/25) |
1.0ul |
PCR Primers:
| k-5'-IO (5'WT) |
5' AGG GTA GGT GTT GGG ATA GC 3' |
| K-3'-1a (3'WT) |
5’ CTC AGT CAT TTT CAG CAG GC 3’ |
| K-3'-neo (3'MUT) |
5’ ACG AGA CTA GTG AGA CTG GC 3’ |
PCR Conditions:
| 1. |
94C |
3:00 |
| 2. |
94C |
1:00 |
| 3. |
60C |
1:00 |
| 4. |
72C |
1:00 |
| 5. |
Goto 2 x33 cycles |
| 6. |
72C |
5:00 |
| 7. |
4C |
0:00 |
| 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:
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