PCR 3 step cycle

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  • PCR
    • 1. Heat to separate strands
      • Heat to 95oC to separate two DNA strands and H Bonds.
    • 2. DNA Primers Added
      • Mixture cool to 53oC to let primers attach to start of each DNA strand.
        • Primer purpose: STOP Rejoining, Seleect are to be COPIED, Signal action of DNA Polymerase
      • Primers easy to identify - radioactive
    • 3.Use of heat-stable DNA polymers - extend primers
      • Mixture heates to 73oC - thermostable DNA Pol. copies each strand.
        • Heat DNA Pol. add free nucleotides to expose strand - attach according to bas pair rules.
          • A-T, G-C

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