Allele and Genotype Lab

Purpose
The purpose of this lab is to get a basic understanding of laboratory procedures and to
learn about PCR (polymerase chain reaction), which is a technology used in Biotech to
copy strands of DNA. The lab also serves to teach us about the importance of
thoroughly reading directions and following the instructions carefully and correctly.
Further, it highlights the importance of labeling things so no DNA gets mixed up.

Hypothesis
If we use PCR to find the frequency of the alu insert in my DNA, then I can figure out
whether I am heterozygous or homozygous.

Materials And Procedures

http://www.babec.org/files/PCR_2012/Alu_PCR_Student_Guide_2012.pdf

Observations
Due to a number of possible errors, our class was unable to figure out individual
genotypes. One possible error could be that we used cubed ice instead of crushed ice. I
also labeled my own DNA with small writing that rubbed off, so I had to guess which one
mine was out of the many that were also rubbed off, which caused much uncertainty.
Another error could have been from improper pipette use. We either could have pipetted
too far or too little, resulting in inconsistent amounts of liquid transferred. Also the PCR
machine could have been on the wrong settings.
Data/Results and Analysis

These are our results. As you can see,

we didn't have much success with this
lab. The line on the left is the ladder.
Since we didn't have any results, I drew
the diagram below. The diagram below
shows where the lines would be if we had
achieved reliable results. You can tell if
someone has the alu segment by looking
to see if they have a fragment at 715 bp
or at 415 bp. If there are only fragments
at 715 bp, then both parents had the alu
segment (+/+): homozygous. If there is
only a 415 bp fragment, neither of them
had the alu segment (-/-): homozygous.
Lastly, if there is both a 715 bp and a 415
bp fragment, then only one parent had the
alu segment in their DNA (+/-):
heterozygous.
Conclusion
Unfortunately we did not get results for this lab, but I did learn some new things, and I
got to practice using laboratory materials such as centrifuges, microfuge tubes, and
micro pipettes. To begin the lab, we first isolated the DNA from cheek cells by swishing
salt water in our mouths for about 30 seconds and then spitting it out in a cup. Then, we
used the PCR (Polymerase Chain Reaction) machine to determine our genotype. A few
days later we went back to the lab and analyzed our results by looking at the gel
electrophoresis under the UV trans illuminator. Because of our lack of results, I have no
general conclusion for this lab because we dont have any evidence on our genotypes. I
would in the future try this lab again because I am eager to figure out if I am
heterozygous or homozygous.