Reaction Components

• According to Integrated DNA Technologies, use a DNA substrate (DNA sample) that is uncontaminated by other DNAs. Other reaction components are water, 10x reaction buffer, MgCl2, dNTPS, forward and reverse primers, and the DNA Polymerase enzyme.
  
  

Considerations

• Polymerase protocol is not appropriate for all DNA samples. DNA templates with a high GC content (Guanine-Cystocene content), low template concentration, or high secondary structure may require more steps than the standard protocol.
  
  

Cycles

• The number of PCR cycles you run is based on the amount of DNA material you have and the amplicons (PCR results) you need to acquire. Twenty-five to 35 cycles is typical for most experiments. (Reference 3)
  
  

Validation

• Validate the resulting amplicons from your PCR reactions using a DNA sequencer if you have access to one. If you don't have access to a sequencer run the final reaction out on agarose gel with an appropriate weight marker to prove the reaction was successful, and that the results are the size you expected.