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Team:SDU-Denmark/Protocols/Primer-PCR
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Protocol for Primer PCR
dNTP mix (10mM)
10ul 100mM dGTP
10ul 100mM dATP
10ul 100mM dTTP
10ul 100mM dCTP
60ul H2O
(Total V=100ul)
PCR reaction
5.0ul Pfx-buffer (÷MgSO4)
1.0ul 50mM MgSO4
1.5ul 10mM dNTP mix
1.5ul forward primer
1.5ul reverse primer
39ul H2O
0.5ul Pfx enzyme (add ju1st before PCR run)
PCR program
| 1. | Start | 94˚C | 2min |
| 2. | Denaturing | 94˚C | 1min |
| 3. | Annealing | 52˚C | 1min |
| 4. | Elongering | 68˚C | 1min |
| 5. | GOTO 2 | rep. 29x | |
| 6. | Slut | 68˚C | 3min |
| 7. | Hold | 4˚C |
