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From 2012.igem.org

7 PCR:

 

7.1 Use TaKaRa Premix Taq Version 2.0. Follow the manufacturer’s protocols.

 

7.2 Set up the following reaction in a PCR tube on ice.

Premix Taq	12.5 μl
Template	0.1 ng - 10 ng
Primer F	0.5 μl
Primer R	0.5 μl
Nuclease-free water	To 25 μl
Total volume	25 μl

7.3 PCR cycle:

94℃ 3min

30cycles

94℃ 30s

55℃ 30s

72℃ 1min

72℃ 5min