Objective:
To determine the frequency of the Alu insert among 5th period Bio/Biotech students.
Materials:
0.9% Saline Solution
Student cheek cells
5% Chelex
Primer Mix
Master Mix ( nucleotides, DNA polymerase, buffer)
TBE buffer
Agarose Gel, tracking dye
DNA stain- "Gel Red"
Positive Control DNA
Base Pair Ladder
Student cheek cells
5% Chelex
Primer Mix
Master Mix ( nucleotides, DNA polymerase, buffer)
TBE buffer
Agarose Gel, tracking dye
DNA stain- "Gel Red"
Positive Control DNA
Base Pair Ladder
Hypothesis:
I think I will be heterozygous which means that I will have an Alu insert on one of my chromosomes.
Procedure:
First, we swished around saline in our mouths to extract our DNA. Then we put the saline solution into a PCR tube. Next we spun it in a micro-centrifuge. Then to draw the cell pellet to the bottom of the tube we racked the tube. After we racked it, we added it to Chelex. Then we inserted a cap lock so that when we heated it overnight the tube didn't pop open from the pressure the heat created. When we came back to the classroom the next day, we took off the cap lock and put the PCR tube in a micro-centrifuge for another minute. We then transferred the supernatant from the Chelex/DNA tube to a new tube, and refrigerated it. After that, we put some of the Chelex/DNA in a small tube, and added Master Mix and Primer Mix. We put it in a thermal cycler and again waited overnight. The next day in class, we spun it in a micro-centrifuge, added loading dye, and put it in a hole in the gel, which was inside of a special gel box. We sent 150 volts through it for about 30 minutes. Then we added a dye to be able to see our DNA which will help us to be able to see what genotype we were.
Result/ Analysis
I was (-.+). I had enough DNA, so my result was very easy to read. I really enjoyed this lab and it was very interesting seeing how biotechnology works. It was a great way to kick off this class.