Team:SDU-Denmark/Protocols/Primer-PCR

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Contents

Protocol for 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 just before PCR run)


PCR program

1. Start 94˚C 2min
2. Denaturing 94˚C 1min
3. Annealing 52˚C 30sek
4. Elongering 70˚C 2min
5. GOTO 2 rep. 29x
6. Slut 70˚C 5min
7. Hold 4˚C
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