Alu PV92 PCR Lab
Purpose
The purpose of this lab was to determine if we were homozygous or heterozygous by finding out if we have Alu in our DNA.
Hypothesis
There are three different kinds of genomes: +/+, -/-, and +/-. This lab will help us determine whether we are heterozygous or homozygous.
Materials
G-Pipet
-Cup
-sharpie
-micro-centrifuge
-chelex
-test tubes
-agarose gel
-master mix
-primer mix
-gel box
-saline solution
-student cheek cells
-TBE buffer
-Cup
-sharpie
-micro-centrifuge
-chelex
-test tubes
-agarose gel
-master mix
-primer mix
-gel box
-saline solution
-student cheek cells
-TBE buffer
Procedure
1. Swish saline solution in your mouth and spit back into a cup
2. Put 1 ml of the saline and cell suspension into microfuge tube
3. Spin microfuge in a centrifuge for one minute
4. Pour off the supernatent but don't lose your pellet cell
5. Rack or flick tube to mix up cells
6. Put cell suspension into tube if Chelex
7. Put cap lock on tube full of cells and Chelex and heat it in a heat block for 10 minutes
8. Put cells and Chelex in a tiny PCR tube and place it in a thermal cycler for 10 minutes
9. After heating, put in centrifuge for 1 minute
10. Withdraw supernatent from Chelex/ DNA into a new microfuge tube
11. Refrigerate isolated DNA
12. Put master mix, primer mix, and DNA into small PCR tube
13. Place tube in thermal cycler
14. Centrifuge tube for 10 seconds
15. Add dye to PCR tube
16. Make gel and let it harden
17. Load DNA mixture into gel
18. Put gel in gel box
19. Turn power on 150 volts for 20-45 minutes
20. Stain gel and photograph it
2. Put 1 ml of the saline and cell suspension into microfuge tube
3. Spin microfuge in a centrifuge for one minute
4. Pour off the supernatent but don't lose your pellet cell
5. Rack or flick tube to mix up cells
6. Put cell suspension into tube if Chelex
7. Put cap lock on tube full of cells and Chelex and heat it in a heat block for 10 minutes
8. Put cells and Chelex in a tiny PCR tube and place it in a thermal cycler for 10 minutes
9. After heating, put in centrifuge for 1 minute
10. Withdraw supernatent from Chelex/ DNA into a new microfuge tube
11. Refrigerate isolated DNA
12. Put master mix, primer mix, and DNA into small PCR tube
13. Place tube in thermal cycler
14. Centrifuge tube for 10 seconds
15. Add dye to PCR tube
16. Make gel and let it harden
17. Load DNA mixture into gel
18. Put gel in gel box
19. Turn power on 150 volts for 20-45 minutes
20. Stain gel and photograph it
Results
By doing this experiment, I learned that I am -/- which means i am homozygous. I am past 400. This lab was very interesting and i really liked learning how to micropipet. At first, this lab made no sense, but i feel comfortable using these tools now. It semmed like a very long lab, but i learned a lot. In this picture, my DNA is in the second column.
Analysis
Compared to all the other students, I was able to retrieve a lot of DNA in the first few steps. I also didn't lose a lot during the other steps, so It was easy to analyze. Mine was most likely brighter than the other ones because the other students probably lost some DNA during the experiment.