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Module 3, Day 1 For Next Time

PCR Cycle

  1. The cycle starts with 94 C for 4 min in order to denature all of the DNA and non-Taq protein in the sample.
  1. Gradually raising the annealing temperature will help in making sure template DNA is annealing to primers instead of itself, because there is more and more template DNA as PCR continues.
  1. It takes more time for the enzyme to polymerize a new strand of DNA than DNA strands to associate/dissociate.
  1. If you were amplifying URA3 on genomic DNA, many things could go wrong.
    • You could amplify the deletion gene if the tails annealed.
    • You could amplify a gene with a similar amino acid sequence.
    • You really just get a lot of things you don't want, because the genome is much more likely than a plasmid to have homologous sequences to your primer.

Writing the Introduction

Module 3 Assignment