IGEM:Groningen/Notebook/iGEM 2011/2011/06/16: Difference between revisions

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(Autocreate 2011/06/16 Entry for IGEM:Groningen/Notebook/iGEM_2011)
 
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==Entry title==
==16-6-11==
* Insert content here...


<br> Obtaining more PCR product:
<br> <br> PCR with taq:
<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
<br> MQ water: 33.5μl
<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
<br>
<br> PCR conditions:
<br> Preheated lid: 111°C
<br> Denaturation: 94°C for 10 min.
<br> Cycle 33×:
<br>  denaturation: 94°C for 30s.
<br>  annealing:    60°C for 30s.
<br>  Extension:    72°C for 2,5 min.
<br> Final extension: 72°C for 10 min.
<br> Store infinite at 4°C
<br>
<br> Analyse on a 1% TBE agarose gel.
<br> Clean up DNA with High Pure PCR Purification Kit
<br>
<br> Team meeting at 4


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Revision as of 02:47, 21 September 2011

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16-6-11


Obtaining more PCR product:

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

Team meeting at 4

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