Team:EPF-Lausanne/Protocols/PCR with Taq Platinium

 

PCR with Taq Platinium Protocol

1. Add the following components to a sterile 0.5-ml microcentrifuge tube. Volumes are for a single 50-μl reaction, and can be scaled as needed. Prepare a master mix of common components for multiple reactions.

*1.0 unit is sufficient for amplifying most targets. In some cases, more enzyme may be required (up to 2.5 units).

2. Cap the tubes, mix, and centrifuge briefly to collect the contents.

3. Incubate tubes in a thermal cycler at 94°C for 30 seconds to 2 minutes to completely denature the template and activate the enzyme.

4. Perform 25–35 cycles of PCR amplification as follows:

5. Maintain the reaction at 4°C after cycling. The samples can be stored at –20°C until use. Analyze the products by agarose gel electrophoresis.