There are at least one pair of primers, including forward primer and reverse primer, which cover the interesting DNA position studied. Designed by forward primers, and then the original DNA was captured on (3'-> 5') or disgusted beaches. The aversion beach part is designed as the backward primer direction, and the original DNA is captured on (5'-> 3') or sensitive beach. Therefore, the primer design must be specific to the original DNA, without generating primer dimer, that is, the two primers have matching pair sequences, and avoid the occurrence of primer self-interaction capture (self-interaction) or needle cycle.
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