Team:SDU-Denmark/Protocols/Primer-PCR

From 2009.igem.org

(Difference between revisions)
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39ul H2O
39ul H2O
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0.5ul Pfx enzyme (add just before PCR run)
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0.5ul Pfx enzyme (add ju1st before PCR run)
   
   
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|  2. || Denaturing || 94˚C || 1min
|  2. || Denaturing || 94˚C || 1min
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|  3. || Annealing || 52˚C || 30sek
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|  3. || Annealing || 52˚C || 1min
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|  4. || Elongering || 70˚C || 2min
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|  4. || Elongering || 68˚C || 1min
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|  5. || GOTO 2 || || rep. 29x             
|  5. || GOTO 2 || || rep. 29x             
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|  6. || Slut || 70˚C || 5min
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|  6. || Slut || 68˚C || 3min
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|  7. || Hold || 4˚C ||  
|  7. || Hold || 4˚C ||  
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[http://www.example.com link title]
 

Revision as of 08:11, 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

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
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