Difference between revisions of "IGEM:Groningen/Notebook/iGEM 2011/2011/06/23"

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(Autocreate 2011/06/23 Entry for IGEM:Groningen/Notebook/iGEM_2011)
 
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==Entry title==
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==23-6-11==
* Insert content here...
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<br>
 
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<br> PCR for PcI, PBADaraC, PlasB and sample of Christoph
 +
<br> <br> PCR with taq:
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<br> 10× taq buffer: 5μl
 +
<br> 10mM dNTPs: 1μl
 +
<br> BB forward primer10μM: 2.5μl
 +
<br> BB reverse primer10μM: 2.5μl
 +
<br> Taq DNA polymerase: 0.5μl
 +
<br> MgCl2 25mM: 4μl
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<br> MQ water: 33.5μl
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<br>
 +
<br> PCR with pfu:
 +
<br> 10× pfu buffer with MgSO4: 5μl
 +
<br> 10mM dNTPs: 1μl
 +
<br> BB forward primer10μM: 1μl
 +
<br> BB reverse primer10μM: 1μl
 +
<br> Pfu DNA polymerase: 1μl
 +
<br> MQ water: 40μl
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<br>
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<br> PCR conditions:
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<br> Preheated lid: 111°C
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<br> Denaturation: 94°C for 10 min.
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<br> Cycle 33×:
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<br>  denaturation: 94°C for 30s.
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<br>  annealing:    60°C for 30s.
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<br>  Extension:    72°C for 2,5 min.
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<br> Final extension: 72°C for 10 min.
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<br> Store infinite at 4°C
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<br>
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<br> Analyse on a 1% TBE agarose gel.
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<br> Clean up DNA with High Pure PCR Purification Kit
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<br>
  
 
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Revision as of 02:35, 21 September 2011

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23-6-11



PCR for PcI, PBADaraC, PlasB and sample of Christoph

PCR with taq:
10× taq buffer: 5μl
10mM dNTPs: 1μl
BB forward primer10μM: 2.5μl
BB reverse primer10μM: 2.5μl
Taq DNA polymerase: 0.5μl
MgCl2 25mM: 4μl
MQ water: 33.5μl

PCR with pfu:
10× pfu buffer with MgSO4: 5μl
10mM dNTPs: 1μl
BB forward primer10μM: 1μl
BB reverse primer10μM: 1μl
Pfu DNA polymerase: 1μl
MQ water: 40μl

PCR conditions:
Preheated lid: 111°C
Denaturation: 94°C for 10 min.
Cycle 33×:
denaturation: 94°C for 30s.
annealing: 60°C for 30s.
Extension: 72°C for 2,5 min.
Final extension: 72°C for 10 min.
Store infinite at 4°C

Analyse on a 1% TBE agarose gel.
Clean up DNA with High Pure PCR Purification Kit