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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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