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  • Polymerase Chain Reaction, PCR, enables researchers to produce millions of

  • copies of a specific DNA sequence in one to two hours.

  • This automated process bypasses the need to use bacteria for amplifying DNA.

  • A PCR reaction starts with a denaturing step.

  • Samples are heated [from] 94 to 96 degrees celsius for 30 seconds or more

  • to denature, or separate in two single strands, the target DNA.

  • Next the temperature is lowered to [between] 50 to 65 degrees celsius for 30 seconds or more.

  • This allows the left and right primers to anneal to their complementary sequences.

  • The primers are designed to bracket the DNA region to be amplified.

  • The temperature is raised to 72 degrees celsius for thirty seconds or more.

  • This allows the taq polymerase to bind each primed site, and extend or synthesize a new DNA strand.

  • The temperature is raised to [between] 94 and 96 degrees celsius, denaturing the target DNA as in cycle one.

  • As before, the temperature cools to [between] 50 and 65 degrees celsius, allowing more primers to anneal.

  • The temperature is again raised to 72 degrees celsius;

  • taq polymerase binds to each primed site and synthesizes the new DNA strand.

  • In subsequent cycles, the process of denaturing, annealing, and extending are

  • repeated to make additional DNA copies.

  • After three cycles, the target sequence defined by the primers begins to accumulate.

  • After thirty cycles, as many as a billion copies of the target sequence are

  • produced from a single starting molecule.

Polymerase Chain Reaction, PCR, enables researchers to produce millions of

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