Molecular Biology 6 - PCR

MolBio 6 Companion Questions

Learning Objective

  • Know the ingredient in PCR
    • Template
    • Forward and Reverse primers
    • dNTPs
    • DNA polymerase
  • Know the general steps in a thermocycler and what they do
    • High 94oC = Denaturing / Melting
    • Low 58oC = Priming / Annealing
    • Medium 72oC = Extension (optimal temp for Taq Polymerase)
  • Know that the annealing temperature depends on primer sequence
    • higher temp increases specificity
    • lower temp for less specificity or for bad primers (short)
  • Be able to design primers for a given target sequence
  • Be able to design primers with added restriction sites
  • Be able to determine the product (size) with the primers you design

Additional Terms

  • Polymerase Chain Reaction
  • Proof-reading