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Team:SDU-Denmark/Protocols/Primer-PCR
From 2009.igem.org
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==PCR reaction== | ==PCR reaction== | ||
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5.0ul Pfx-buffer (÷MgSO4) | 5.0ul Pfx-buffer (÷MgSO4) | ||
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1.5ul reverse primer | 1.5ul reverse primer | ||
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0.5ul Pfx enzyme (add ju1st before PCR run) | 0.5ul Pfx enzyme (add ju1st before PCR run) | ||
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==PCR program== | ==PCR program== | ||
Latest revision as of 08:12, 20 August 2009
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Contents |
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
39ul H2O
5.0ul Pfx-buffer (÷MgSO4)
1.0ul 50mM MgSO4
1.5ul 10mM dNTP mix
1.5ul forward primer
1.5ul reverse primer
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 |
