Biomedical Innovations

Lesson 4.1.2 PCR

11 PA Points

Part I: PCR Basics

Go through the powerpoint presentation and answer the following questions.

1. Explain the meaning of the words polymerase and chain reaction. 1 point
Polymerase = The enzyme polymerase makes polymers of molecules in our case DNA.
Chain Reaction = A chemical reaction that progresses in an exponential way.

2. What is the purpose of PCR? 1 point

PCR is an efficient and cost effective way to copy or amplify small segments of DNA or
RNA.

3. Who invented PCR and what year? When? 1 point

PCR was invented by Dr. Kary Mullis in 1984.

4. List the three uses of PCR? 1 point

Diagnose infections to identify pathogens using a minimal amount of sample, such as
blood or tissue.
Crime scene investigations
Genetic research
Biotechnology .
Paternity testing
Evolutionary relationships

DNA FingerPrints, Medical Diagnostic, Evolutionary Relationships

5. Explain the role of each component of PCR in detail in the table below. Use this website
for information. 1.5 point
https://www.thermofisher.com/us/en/home/life-science/cloning/cloning-learning-
center/invitrogen-school-of-molecular-biology/pcr-education/pcr-reagents-enzymes/
pcr-component-considerations.html
Component Role in PCR

DNA They can generate new strands of DNA using a DNA template and
primers, and they are heat resistant.

Taq polymerase
Primers 2 sets (forward and reverse), synthetically produced. Adds
nucleotides to the 3' end.

Nucleotides (dNTP)
dNTP expands the growing DNA strand. It binds with the DNA
strand by hydrogen bonds.

MgCl2 (Mg ion) The role of MgCl2 (Mg ion) is to stabilize the binding of DNA
polymerase to the template DNA.

Buffer Buffer provide a suitable chemical environment for activity of DNA

polymerase.

6. On what end of the DNA does Taq polymerase add nucleotides? 1 point
3’ ends.

7. Why is Taq DNA polymerase used in PCR and not the polymerase used by mammalian
cells to synthesize cellular DNA? 1 point
Taq polymerase is used in PCR because it is heat-resistant and has a high error rate,
making it suitable for the denaturation step and repeated reactions.

8. What is a thermocycler? 1 point

A thermocycler carries out PCR.

9. The three steps of PCR are listed in the table below. Describe in detail what happens in
each step, including the temperatures and components involved. 1.5 points
Name of Step Description

Denaturation The tube containing the sample DNA is

heated to more than 90 °C (194° F), which
separates the double-stranded DNA into two
separate strands.

Annealing Binding DNA Primers to the DNA Sequence.

Extension DNA Sequence is Duplicated

10. After 15 cycles of PCR how many copies of DNA would there be?
32,768 copies of DNA.

I got this from 2^15.