Polymerase Chain Reaction Laboratory
Abstract:
In this lab, we analyzed DNA from our cheek cells to determine whether or not we had the Alu sequence in our DNA. The Alu segment can help determine where a person's ancestors our from, but it does not affect human traits, so a DNA test must be performed to tell if someone has it.
In this lab, we analyzed DNA from our cheek cells to determine whether or not we had the Alu sequence in our DNA. The Alu segment can help determine where a person's ancestors our from, but it does not affect human traits, so a DNA test must be performed to tell if someone has it.
Purpose:
The purpose of this lab was to learn about the polymerase chain reaction (PCR), to learn about laboratory procedures, and to determine whether or not our genes contain the Alu segment. PCR is a very important technique to learn as a bio-technician, as it is the most effective way to analyze DNA.
The purpose of this lab was to learn about the polymerase chain reaction (PCR), to learn about laboratory procedures, and to determine whether or not our genes contain the Alu segment. PCR is a very important technique to learn as a bio-technician, as it is the most effective way to analyze DNA.
Hypothesis:
I do not believe that I will have the Alu repeat in my genes, seeing as it is less common in European descendants. (A Sajantila, University of Helsinki)
I do not believe that I will have the Alu repeat in my genes, seeing as it is less common in European descendants. (A Sajantila, University of Helsinki)
Materials and Procedure:
The full procedure and all materials used can be found on a PDF document at the web address below.
http://academic.regis.edu/tnakamur/Alu_student.pdf
Materials:
Centrifuge Cheek Cells
Chelex Master Mix
Primer Mix Electric Scale
TBE Buffer Gel Casting Tray
Tape Agarose Gel
DNA Stain Solution
Procedure:
We started this lab by swirling a saline solution in our mouths. Then we expectorated(spit) into a cup and mixed it. We then labeled a centrifuge tube with our initials and extracted one milliliter of the DNA into the tube. Next, we put the mixture into the centrifuge and spun it for one minute. After that, we removed the top of the solution, mixed it, and added 50 microliters of Chelex. Then we heated it with a heat block for about ten minutes. We then released the pressure in the tube and put it back on the centrifuge for another minute of spinning. Following this, we removed 50 microliters and placed it into a new tube, and refrigerated it for a night. The next day, we added 20 micrometers of Master mix into the tube, then took 10 microliters of our DNA for the tube with Chelex and placed it into the new tube. Finally, we heated up the solution on the heat block.
Results:
I did not have the Alu segment in my DNA, making me homozygous and --. Interestingly, it seemed like there was an even split between homozygous and heterozygous students.
I did not have the Alu segment in my DNA, making me homozygous and --. Interestingly, it seemed like there was an even split between homozygous and heterozygous students.
Analysis:
Neither my mother nor my father has the Alu repeat in their DNA. If one of them had it, there would have been a 415 bp fragment, and an 715 bp fragment. If both of them had it, there would only be a fragment at 715 bp. In my case, I only had a fragment at 415 bp, meaning neither of my parents has the Alu repeat.
Neither my mother nor my father has the Alu repeat in their DNA. If one of them had it, there would have been a 415 bp fragment, and an 715 bp fragment. If both of them had it, there would only be a fragment at 715 bp. In my case, I only had a fragment at 415 bp, meaning neither of my parents has the Alu repeat.
Conclusion:
While learning about my DNA was interesting, I believe that the most important part of the lab was learning about proper biotech procedures. The class made a mistake in the first part of the lab, and unfortunately had to start over. Although this was a bummer, we learned that this is frequent in biotech experiments, and it is important not to give up on the lab. We also got to practice using laboratory materials such as centrifuges, microfuge tubes, and micro pipets. Overall, this was a great way to learn about safe lab practices, proper use of equipment (balancing the centrifuge), and PCR.
While learning about my DNA was interesting, I believe that the most important part of the lab was learning about proper biotech procedures. The class made a mistake in the first part of the lab, and unfortunately had to start over. Although this was a bummer, we learned that this is frequent in biotech experiments, and it is important not to give up on the lab. We also got to practice using laboratory materials such as centrifuges, microfuge tubes, and micro pipets. Overall, this was a great way to learn about safe lab practices, proper use of equipment (balancing the centrifuge), and PCR.