The ALU PCR Lab was an interesting way to start off the year. In the lab, we analyzed our cheek DNA to determine if we had the ALU segment in our DNA or not. The ALU segment has no effects on human traits, so it's impossible to determine if one has the segment unless a DNA test is administered.
Purpose
The reason we did this lab was to get a basic understanding of laboratory procedures and to learn about the polymerase chain reaction, or PCR, a technology used in Biotech fields to amplify and copy strands of DNA. The lab also taught us about the importance of reading and following instructions correctly and carefully, and the importance of labeling things so you do not forget what they are.
Hypothesis
I hypothesize that I will have the gene from one of my parents, but not the other. This is based off a University of Helsinki study by A. Sajantila that found that ALU is less common in European descendants. Since my mother is Chinese and my father a mix of northern European, it makes sense that I would have half of it.
Materials
- Micropipettes (p20 and p200), tips, and tubes
- Microcentrifuge
- Heat block
- Saline solution
- Human cheek cells
- Master mix
- Primer mix
- Chelex
- Agarose gel
- DNA stain solution
Procedure
We began by swirling saline solution in our mouths. We then spit it out into a cup (now containing our DNA) and swirled it around to mix it. We then labeled a microcentrifuge tube with our initials and pipetted 1000 microliters of the DNA into the tube. After that, we spun the mixture in a microcentrifuge for one minute. We then poured off some of the top solution. After that, we remixed it and added 50 microliters of the mix into a tube of Chelex. We then heated it on the heat block for 10 minutes. After that, we released the pressure in the tube and respun it on the microcentrifuge for another minute. After that, we took out 50 microliters and placed it into a new tube, which we refrigerated.
After a night of refrigerating, we added 20 microliters of Master mix into the tube. After that, we added the same amount of Primer mix into the same tube. We then took 10 microliters of our DNA from Chelex tube and placed it into the new tube. We then heated up the solution on the heat block.
Results
It turns out that I am -/- (minus - minus). This means that I have not inherited any of the ALU segment from my parents.
Analysis
Neither of my parents have the Alu repeat in their DNA. You can tell if someone has the ALU segment by looking to see if they have a fragment at 715bp or at 415bp. If there are only fragments at 715bp, then both parents had the ALU segment. If there is only a 415bp fragment, neither of them had it. However, if there is both a 715bp and a 415bp fragment, then only one parent had the ALU segment in their DNA. I only had a 415bp fragment, indicating that neither of my parents have the ALU segment, and neither do I.
Conclusion
Even though learning about our own DNA was very interesting, I feel that the most important part of this lab was learning about the basic procedures and protocols in a biotechnology laboratory. A good example of this was how the class messed up one of the parts in the beginning, forcing us to restart. At first glance this only seems like an annoyance, but upon further inspection, it really helped teach us how not everything goes right on the first try, and how true scientists not only have to be curious and intelligent, but also determined and persistent. The lab also taught us a great deal about pipetting, centrifuges, microfuge tubes, and other standard laboratory materials.
Purpose
The reason we did this lab was to get a basic understanding of laboratory procedures and to learn about the polymerase chain reaction, or PCR, a technology used in Biotech fields to amplify and copy strands of DNA. The lab also taught us about the importance of reading and following instructions correctly and carefully, and the importance of labeling things so you do not forget what they are.
Hypothesis
I hypothesize that I will have the gene from one of my parents, but not the other. This is based off a University of Helsinki study by A. Sajantila that found that ALU is less common in European descendants. Since my mother is Chinese and my father a mix of northern European, it makes sense that I would have half of it.
Materials
- Micropipettes (p20 and p200), tips, and tubes
- Microcentrifuge
- Heat block
- Saline solution
- Human cheek cells
- Master mix
- Primer mix
- Chelex
- Agarose gel
- DNA stain solution
Procedure
We began by swirling saline solution in our mouths. We then spit it out into a cup (now containing our DNA) and swirled it around to mix it. We then labeled a microcentrifuge tube with our initials and pipetted 1000 microliters of the DNA into the tube. After that, we spun the mixture in a microcentrifuge for one minute. We then poured off some of the top solution. After that, we remixed it and added 50 microliters of the mix into a tube of Chelex. We then heated it on the heat block for 10 minutes. After that, we released the pressure in the tube and respun it on the microcentrifuge for another minute. After that, we took out 50 microliters and placed it into a new tube, which we refrigerated.
After a night of refrigerating, we added 20 microliters of Master mix into the tube. After that, we added the same amount of Primer mix into the same tube. We then took 10 microliters of our DNA from Chelex tube and placed it into the new tube. We then heated up the solution on the heat block.
Results
It turns out that I am -/- (minus - minus). This means that I have not inherited any of the ALU segment from my parents.
Analysis
Neither of my parents have the Alu repeat in their DNA. You can tell if someone has the ALU segment by looking to see if they have a fragment at 715bp or at 415bp. If there are only fragments at 715bp, then both parents had the ALU segment. If there is only a 415bp fragment, neither of them had it. However, if there is both a 715bp and a 415bp fragment, then only one parent had the ALU segment in their DNA. I only had a 415bp fragment, indicating that neither of my parents have the ALU segment, and neither do I.
Conclusion
Even though learning about our own DNA was very interesting, I feel that the most important part of this lab was learning about the basic procedures and protocols in a biotechnology laboratory. A good example of this was how the class messed up one of the parts in the beginning, forcing us to restart. At first glance this only seems like an annoyance, but upon further inspection, it really helped teach us how not everything goes right on the first try, and how true scientists not only have to be curious and intelligent, but also determined and persistent. The lab also taught us a great deal about pipetting, centrifuges, microfuge tubes, and other standard laboratory materials.