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PCR Techniques · Pathology

How does 'Hot-Start' PCR improve the specificity of DNA amplification?

  1. By increasing the temperature of the extension step to 80°C
  2. By using a heat-activated polymerase inactive at room temperature
  3. By adding chilled primers directly to the tube during the first cycle
  4. By performing the reaction in a vacuum-sealed chamber
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Correct answer: By using a heat-activated polymerase inactive at room temperature

Hot-start PCR prevents the formation of non-specific products and primer dimers during the initial setup at room temperature. The DNA polymerase is typically inhibited by an antibody or chemical modification that is only released upon heating.

Difficulty: Medium Question 8 of 20

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