RT-PCR · Pathology

Why is the use of 'DEPC-treated water' important during the preparation of RT-PCR reagents?

  1. It inhibits DNA polymerase to ensure specificity
  2. It inactivates RNases to prevent RNA degradation
  3. It provides the necessary ions for primer annealing
  4. It acts as a fluorescent dye for detection
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Correct answer: It inactivates RNases to prevent RNA degradation

RNases are ubiquitous and very stable enzymes that degrade RNA. Diethylpyrocarbonate (DEPC) is a chemical that covalently modifies and inactivates these enzymes, ensuring the RNA template remains intact during the assay setup.

Difficulty: Medium Question 15 of 20

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