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PCR Polymerase Chain Reaction a tool to generate a large amount of a gene of interest DNA DNA-sequence in a plasmid Plasmid Restriction of plasmid releases the DNA sequence of interest Polymerase Chain Reaction First cDNA strand synthesizes only the gene of interest by using Forward and Reverse primers (specific small pieces of DNA) with the help of the Reverse primer Second cDNA strand enzyme Polymerase. All cycles follow a strict order of timed temperatures. Gene of Interest Forward primer