IGEM:Groningen/Notebook/iGEM 2011/2011/06/10

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(Autocreate 2011/06/10 Entry for IGEM:Groningen/Notebook/iGEM_2011)
Current revision (04:23, 21 September 2011) (view source)
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==Entry title==
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==10-06-11==
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* Insert content here...
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<br>
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<br> Colony PCR of transformants!
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<br> Taq 10× buffer: 40μl
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<br> dNTPs 10mM: 8μl
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<br> Forward Biobrick primer: 8μl
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<br> Reverse Biobrick primer: 8μl
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<br> Taq DNA polymerase: 2μl
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<br> MQ water: 334μl
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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>
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<br> Asked info in team about why some PCR's fail: next time use for taq buffer not the buffer with KCI, but NH4SO4
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<br>
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<br>
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10-06-11



Colony PCR of transformants!
Taq 10× buffer: 40μl
dNTPs 10mM: 8μl
Forward Biobrick primer: 8μl
Reverse Biobrick primer: 8μl
Taq DNA polymerase: 2μl
MQ water: 334μl

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.

Asked info in team about why some PCR's fail: next time use for taq buffer not the buffer with KCI, but NH4SO4

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