PCR

Question 1

Denaturation

Answer 1

DNA heated to 94% which allows dsDNA to separate into two ssDNA strands

Question 2

Annealing

Answer 2

Cooled to 55-60 degrees, primers bind to each ssDNA strand

Question 3

Elongation

Answer 3

Heated to 72 degrees, Taq polymerase (Pfu for higher specificity) brings free dNTPs to the ssDNA extending it in a 5’ to 3’ direction from the primer

Question 4

Taq vs Pfu

Answer 4

Taq cheaper and quicker, Pfu more specific with less error rates.

Taq - Thermus aquaticus - derived from bacteria that live in hot springs so they can withstand high temperatures

Pfu - Pyrococcus furiosus