Module 3, Day 1 For Next Time
- The cycle starts with 94 C for 4 min in order to denature all of the DNA and non-Taq protein in the sample.
- 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.
- It takes more time for the enzyme to polymerize a new strand of DNA than DNA strands to associate/dissociate.
- 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.