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Primer sequences are determined from known sequence since there must be a match to the region of DNA to be amplified. PCR amplification requires 2 primers that determine the region of sequence amplified in the forward and reverse direction.ikewise, the reverse primer is designed from the complimentary strand. PCR is exponential amplification in which the newly generated PCR fragment from one cycle also acts as a template for the next cycle (Figure 1A). Amplification of DNA for Sanger sequencing differs from PCR in that a single primer is used.
Primer Design Parameters for Multiplex PCR. Design of specific primer sets is essential for a successful multiplex reaction. The important primer design considerations described below are a key to specific amplification with high yield.