Purpose: For this lab, we studied DNA to learn about its origin and/or its history, how it’s still evolving, and to gain overall lab experience.
Hypothesis: If we used PCR to analyze the DNA, then I could’ve used it to learn the history of my DNA, the amount of Alu repeats, and from where my DNA originated.
Procedure: We worked in accordance with BABEC(Bay Area Bioscience Education Consortium) Alu PCR, 2017
Data: We used 2 percent agarose gel that we ran at 150 U for 20 minutes and stained using gelred for 72 hours. Lane A1 and 2A have 100 bp ladder. Lane 1 B-H and B-18 have 20 mL of a 50 mL DNA/10 mL loading dye solution. Lane B section 3 contains my DNA sample. I didn’t get any results, making my data inconclusive.
When defining genotype, it can be best defined as the type of alleles an organism holds within their DNA. There are three different variations: +/+, +/-, -/-. The calculations show an ideal simulation of what each student would be in the classroom, with a total of 37 students. 15 being +/+, 10 being +/-, and 12 being -/-. To find the total number of alleles, we have to take the total number of students and multiply it by 2 because every student has two alleles. This brings us to a total of 74 alleles. Now, to find the total number of positive alleles, we take the 15+/+ and multiply that by 2, giving us 30 positives. Then, we take the 10 from the 10+/- and add that to 30, giving us a total of 40 “+” alleles. To find the negative alleles, we take the 10 from the 10+/- and add that to the 12-/- students. Because you multiply 12 by 2 and get 24, you add that to 10 for a grand total of 34 “-” alleles. Errors: There were many things that could’ve contributed to my failure to get results. All of which were human errors like:
I could’ve not cleaned my mouth well enough with the salt water .
I could’ve not spit enough DNA for it to have lasted.
I might’ve spun it too much.
The pipette measurements could have been wrong.
I might’ve centrifuged it for too long .
Improvements For Next Time:
Using a larger sample of DNA could help because there would be a lower chance of it drying out.
Having longer to do it next time because we didn’t accurately do everything, so if we had more time we’d be able to be more precise.
How we manage our time is a big factor, focusing more would’ve meant that we would’ve done things more accurately.
Conclusion: I discovered that following through with a lab is actually a lot harder than I had previously imagined. It was difficult to follow every single step closely and make sure that I executed the steps in the appropriate fashion. I thought by using PCR, I’d be able to trace the history of my DNA and see its origin. Sadly, no luck, it didn’t show up in the gel. Mistakes were definitely made, such as centrifuging it for too long or not spitting enough DNA to start with. I remember putting the DNA in the centrifuge one more time than I needed on accident. This most likely ruined my sample. It was also really important to put a certain amount into the gel during the final stages. I remember not putting in the exact amount, this also contributed to me not getting results. In conclusion, this was a very intricate and complicated lab, and my lack of attention was the definite reason that I didn’t end up getting results.
REFLECTION:
This was definitely one of the hardest projects I've had to do. It was the most intricate and you needed to really pay attention if you wanted to understand it. Every little step had to be done and done correctly if you wanted to get results. Obviously, since the majority of the class didn't get results, we didn't do every step right. I learned that I might not be as smart as I thought I was, this was the first time I was lost in a project. Even though it was my fault for not paying enough attention, it was still a very hard and complicated project. I think a lot of my teammates learned the same thing. It was also difficult because even though we were in groups, it was mostly individual because everyone had to do their own thing. In conclusion, I really need to give my all to make sure I can get the most out of a project.