PCR Techniques · Pathology

Which of the following modifications is used in Allele-Specific PCR to detect a Single Nucleotide Polymorphism (SNP)?

  1. Designing primers where the 3' terminal base matches the specific SNP
  2. Using RNA instead of DNA as the template
  3. Increasing the PCR extension time from the standard 1 minute to 10 minutes
  4. Adding radioactive isotopes to the master mix
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Correct answer: Designing primers where the 3' terminal base matches the specific SNP

DNA polymerase requires a perfectly matched 3' end to initiate extension. By placing the SNP site at the 3' end of the primer, amplification will only occur if the specific allele is present in the sample.

Difficulty: Medium Question 15 of 20

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