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Team:SDU-Denmark/Protocols/Primer-PCR
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[[Team:SDU-Denmark|Home]] | [[Team:SDU-Denmark/Background|Background]] | [[Team:SDU-Denmark/Project|Project]] | [[Team:SDU-Denmark/Parts|Parts]] | [[Team:SDU-Denmark/Team|Team]] | [[Team:SDU-Denmark|Home]] | [[Team:SDU-Denmark/Background|Background]] | [[Team:SDU-Denmark/Project|Project]] | [[Team:SDU-Denmark/Parts|Parts]] | [[Team:SDU-Denmark/Team|Team]] | ||
[[Team:SDU-Denmark/Diary|Diary]] | [[Team:SDU-Denmark/Protocols|Protocols]] | [[Team:SDU-Denmark/Downloads|Downloads]] | [[Team:SDU-Denmark/Brainstorm|Brainstorm]] | [[Team:SDU-Denmark/Diary|Diary]] | [[Team:SDU-Denmark/Protocols|Protocols]] | [[Team:SDU-Denmark/Downloads|Downloads]] | [[Team:SDU-Denmark/Brainstorm|Brainstorm]] | ||
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=Protocol for Primer PCR= | =Protocol for Primer PCR= | ||
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==PCR reaction== | ==PCR reaction== | ||
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| + | 39ul H2O | ||
5.0ul Pfx-buffer (÷MgSO4) | 5.0ul Pfx-buffer (÷MgSO4) | ||
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1.5ul reverse primer | 1.5ul reverse primer | ||
| - | + | 0.5ul Pfx enzyme (add ju1st before PCR run) | |
| - | + | ||
| - | 0.5ul Pfx enzyme (add | + | |
| - | + | ||
| - | + | ||
==PCR program== | ==PCR program== | ||
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| 2. || Denaturing || 94˚C || 1min | | 2. || Denaturing || 94˚C || 1min | ||
|- | |- | ||
| - | | 3. || Annealing || 52˚C || | + | | 3. || Annealing || 52˚C || 1min |
|- | |- | ||
| - | | 4. || Elongering || | + | | 4. || Elongering || 68˚C || 1min |
|- | |- | ||
| 5. || GOTO 2 || || rep. 29x | | 5. || GOTO 2 || || rep. 29x | ||
|- | |- | ||
| - | | 6. || Slut || | + | | 6. || Slut || 68˚C || 3min |
|- | |- | ||
| 7. || Hold || 4˚C || | | 7. || Hold || 4˚C || | ||
|} | |} | ||
Latest revision as of 08:12, 20 August 2009
Home | Background | Project | Parts | Team
Diary | Protocols | Downloads | Brainstorm
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 |
