PCR Lab Report

Oisín Ó Dónaill

PCR, or polymerase chain reaction, is a technique used to create multiple

samples of a DNA segment for further testing. Due to the large amount of
sample DNA required for genetic and molecular analysis, PCR is vital to this
process as it provides this vast quantity of DNA for analysis (1). This practical
aims to use PCR and a strand of hair recovered at the scene to provide
sufficient genetic evidence to prosecute a suspect in a burglary case.

Source: https://commons.wikimedia.org/wiki/File:Polymerase_chain_reaction.svg

PCR is a technique discovered in 1983 by American biochemist Kary Mullis, and is used to
generate up to millions of copies of a section of DNA from a small amount of said DNA. PCR
has 3 main stages; 1: Denaturing, where the double-stranded template DNA is heated to 94-
95°C to separate it into two separate strands; 2: Annealing, where the temperature is
lowered to between 50-56°C to allow DNA primers to attach to the template DNA; and 3:
Extending, when the temperature is raised to 72°C to allow the Taq polymerase enzyme to
form a new strand of DNA. (2)

Method
1: Gather the five samples of labelled DNA into a tube rack, which already contain the Taq
polymerase and raw nucleotide building blocks used in the PCR process.
2: Add the appropriate primer to a pipette and add to the DNA sample. Repeat this process
for all DNA samples and then again with the second primer.
3: Place the samples into the PCR machine and close the lid. Press the green arrow to start
the PCR process. Wait until the screen reads “Complete” and then open the lid of the
machine. Return the tubes to the tube rack, maintaining the same order as before they
were placed into the machine.
4: Using a pipette, take a sample of the now amplified genes from the tube and place into
the appropriate well of the gel electrophoresis machine. Repeat this with all other samples.
5: Once the samples have been placed in their wells, attach the diodes to the machine and
press the power button on the machine’s power supply. Then press the grey button that
reads “Off” and it will change to “On” and the process will begin.
6: Using a UV light, analyse the distance travelled by the fragments and pay attention to the
brightest band in each lane where the fragments of the targeted gene accumulate. Record
all results in your lab book.

Results

Through use of PCR and gel electrophoresis, we see that suspect 3, in lane 5, matches the crime
scene DNA in lane 2. This tells us that suspect 3 is the criminal. We can see that the amplified genes
have accumulated in the same spots between both samples thanks to the electrophoresis which tells
us they are genetically identical to each other.
 References
1. Polymerase Chain Reaction (PCR) Fact Sheet [Internet]. Genome.gov. 2022 [cited 3 May 2022].
Available from: https://www.genome.gov/about-genomics/fact-sheets/Polymerase-Chain-Reaction-
Fact-Sheet

2. What is PCR (polymerase chain reaction)? [Internet]. Yourgenome.org. 2022 [cited 4 May 2022].
Available from: https://www.yourgenome.org/facts/what-is-pcr-polymerase-chain-reaction