Hannah Pennington

Informative Speech Outline

Topic: Polymerase Chain Reaction (PCR) and its real life applications
Specific Purpose: My goal is to inform the audience of the mechanism and process of a procedure, PCR,
that is used daily in a variety of labs and the impact it has on their lives.
Thesis: PCR was developed by Kary Mullis, who utilized the mechanisms of our cells and applied a three-
stage model for the replication of DNA in-vitro, which is used in a variety of fields.
Introduction:
I. Attention Material: Helena Greenwoods murder case was reopened and solved fifteen years
following her death due to the ability of PCR to amplify the DNA in skin fragments found under
her nails at the time of her death.
II. Credibility Material: This summer, I worked in a cancer research laboratory where I used PCR
nearly every day.
III. Preview: PCR was developed by Kary Mullis, who utilized the mechanisms of our cells and
applied a three-stage model for the replication of DNA in-vitro, which is used in a variety of
fields.
Body:
I. First Main Point: DNA is the reason that we can function and why we are all different. PCR can
replicate small amounts of DNA and create billions of copies quickly and efficiently.
A. Subpoint: DNA is composed of four different bases. The ordering of these bases is what
codes for every part of our life.
B. Subpoint: PCR can be used to amplify a certain region of DNA to any desired amount.
C. Subpoint: DNA is double stranded. The second strand of DNA is opposite to the first.
Transition:
This nature of the DNA is what allows the replication of the strands in the cells, and is
what we take advantage of to replicate the cells our self.
II. Second Main Point: PCR was invented by Kary Mullis in 1983.
A. Subpoint: The concept of using polymerase, the molecule which actually replicates the
DNA, in-vitro was not new
B. Subpoint: What was new was the idea of using the products for the next step
C. Subpoint: The isolation of Taq polymerase was a timely discovery that further eased the
process of PCR
i. PCR originally used DNA polymerase from E. coli, which often denatured at
the high temperatures required for denaturation of DNA
Transition:
Mulliss work in understanding the cyclic ability of replication led to the development of
a general three stage process that serves as the basis of PCR.
III. Third Main Point: PCR mimics the mechanisms of DNA replication inside your body to do the
same in the lab.
A. Subpoint: Denaturation separates the DNA into two single strands.
Hannah Pennington
Informative Speech Outline
i. Typically 94 98 C for 20-30 seconds
ii. The high temperature disrupts the hydrogen bonds between the base pairs.
B. Subpoint: Annealing gives a start point for replication.
i. The temperature is lowered to 50 - 65 C for 20 40 seconds.
ii. A primer, picked specifically for whatever fragment you are trying to
replicate, binds to each single strand at the target.
C. Subpoint: Extension replicates the DNA two create two complete strands from one.
i. A special polymerase, Taq polymerase, which functions only at a specific
temperature range (75 - 80 C), synthesizes new DNA from DNTPs in the
solution.
ii. Can synthesize up to 1000 bpm, time is dependent upon this
Transition:
By cycling through these three phases numerable times (typically 20-40 times), the DNA
can be amplified to quantities usable for testing.
IV. Four Main Point: PCR is used in many different places.
A. Subpoint: Selective DNA Isolation utilizes PCR to amplify only a target sequence for uses
such as determining relationships
B. Subpoint: PCR is useful because it amplifies the amount of DNA, providing enough
sample for testing from very small amounts.
C. Subpoint: PCR can be used to discriminate DNA for diagnosis of disease.

Conclusion:
I. Summary: The discovery of PCR was due to the knowledge of DNA replication in the cell.
Mullis applied this and exponential growth to create the three stage cycle that is used across
the world in labs daily.
II. Concluding Remarks: If you are ever in danger of being murdered, it may be in your best
interest to get a little skin, so that the investigators may extract and amplify that DNA to
prosecute your killer.