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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.
To make primers of the correct sequence that will bind to the template DNA, it is necessary to know a little bit of the template sequence, on either side of the region of DNA you wish to amplify. DNA polymerase and dNTPs are commercially available from biotechnology supply companies.he template is the DNA that contains the target you want to amplify. The primers are synthetic DNA molecules whose sequence matches a region flanking the target you are trying to amplify. It is possible nowadays, to chemically synthesize DNA molecules of any given base sequence, to use as primers.