PCR Reactions

Question 1

Step 1) Denaturation

Answer 1

Is heated to 94 C which denatures the dsDNA to ssDNA

Question 2

Step 2) Annealing

Answer 2

The reaction is cooled to 55 C to allow primers to anneal to each template strand

Question 3

Step 3) Extension

Answer 3

PCR reaction is heated to 72 C.
Taq polymerase synthesizes a strand of DNA complementary to the template strands by extending from the 3’ ends of the primers

Question 4

What 2 primers are used in the PCR reaction?

Answer 4

PGloAmprF (forward) which anneals to the 5’ end of the coding region
PGloAmpR (reverse) which anneals to the 3’ end of the coding region

Question 5

What is PCR?

Answer 5

An in vitro technique that can rapidly amplify a target DNA sequence of interest