Alu Pcr lab write-up
Overview/Purpose:
In this lab we used the method known as Polymerase Chain Reaction, or PCR for short, (Cause who wants to say Polymerase Chain Reaction!) to determine whether or not we have the Alu strand in our DNA. The purpose was to determine the frequency of the Alu insert in the STEM Biology class using PCR.
Materials:
In this lab we used:
- Chelex
- Cheek Cells
- Master Mix (Nucleotides, DNA, Polymerase)
- Primer Mix
- TBE buffer, Agarose Gel
- DNA stain solution
- Microcentrifuge
- 20/200/1000 mL micropipets
- Goggles
- Epindorf tubes
- Heat block
- Saline Solution (0.9%)
Procedure:
Here are the steps we followed to perform this experiment:
- First we swirled saline in our mouths to extract the cheek cells
- We then extracted 1mL into a separate epindorf tube
- We spun this in a micro-centrifuge for a minute to pellet the cells
- We poured off some excess liquid, then racked the rest
- We took 50 microliters of chelex and put it in our mix
- We put the tubes in a heat block for 10 minutes
- We extracted 50 microliters of this and separated this
- We put 20 microliters each of the master and primer mixes into the separated bit
- We dyed it and put in a gel to compare its base pairs
Results:
When we got our results back, it turned out that I had heterozygous results. This means that I had one band matching 400 and 715 on the base pair ladder. You can see this, kinda, in the picture below. My results are second from the right.
Analysis/Conclusion:
This project taught me many useful skills. I learned how to use the pipettes and extract fluids, how to balance the centrifuge, and much more. I also learned that I was heterozygous (+-), with one of my parents having the Alu gene and the other not. This was a useful lab to learn about the scientific method, and also a nifty fact about myself.