IGEM:Groningen/Notebook/iGEM 2011/2011/06/03

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03-06-11


Colony PCR of transformants
Mastermix:
Taq 10× buffer: 116μl
dNTPs 10mM: 23.2μl
MgCl2: 69.6μl
Forward Biobrick primer: 23.2μl
Reverse Biobrick primer: 23.2μl
Taq DNA polymerase: 5.8μl
MQ water: 899μl

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.

Working on Human Practices.

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