I Got It From My Mama: Mitochondrial DNA Lab

Introduction
a. Background- In this lab, we will be testing mitochondrial DNA to see if an individual carries a gene for a disease. We receive our mitochondrial DNA from our moms. In order to do so, one must test the genome, which is our hereditary code. We will be extracting our own mitochondrial DNA and mass producing in through PCR (Polymerase Chain Reaction). The necessary components for PCR are primers, nucleotides, DNA polymerase, and the existing DNA template to be replicated. PCR is beneficial because it can replicate large amounts of DNA that were formerly not able to be replicated. After replicating the gene, we will run the DNA through a gel, which will show us if we carry the gene for the disease. 
b. Purpose- This procedure is important because it could be used to detect if one carries the genes for a disease before symptoms actually occur. This could mean lives and large amounts of money being saved by early detection of diseases. It is also important because it could help us determine if we got a disease-causing gene from our mothers. 
c. Summary- On Day 1 of the lab, we will be extracting our mitochondrial DNA. first we will collect DNA from our cheek cells and break open the cell membranes. We will also add instagene matrix to kill DNAse, which could harm the DNA. On Day 2, we will be using PCR to replicate the small amount of DNA that we extracted. Primers used in PCR target the "disease gene" in the genome. On Day 3, we use gel electrophoresis to diagram the results of the test.
d. Hypothesis- I hypothesize that one of us will have the "disease gene".

Procedure
On day 1, we rinsed our mouths vigorously with saline for 30 seconds. We transfered 1 ml of the saline rinse into a micro test tube containing 200 microliters of InstaGene matrix. We then centrifuged the tubes for 2 minutes, and saw a pellet of cell appear at the bottom of the tube. After pelleting the cells, we poured off the saline and re suspended the pellet by vortexing the tube. Next we micropipetted 20 microliters of of cells into the tube containing InstaGene, and then again vortexed the tube. Then we incubated the tube at 56 degrees Celsius for ten minutes, but at 5 minutes we vortexed the tubes before returning them back to the bath. After, we placed the tubes in a boiling water bath for 5 minutes. Then, we vortexed the contents for 5 minutes and refrigerated the tubes overnight. On day 2, we centrifuged the tubes for 2 minutes. Then we transferred 20 microliters of the DNA template into the PCR tube being sure to not transfer any matrix beads. We also added 20 microliters of the master mix into the PCR tube. Finally we placed the tubes into the thermal cycler. On day 3, we obtained our PCR tubes and centrifuged them for 3 seconds. Then we added 10 microliters of loading dye into the PCR tube and mixed it gently. After we pipeted 10 microliters of the marker into lane 1 of the gel. In lanes 2, 3, 4, and 5, we each placed 20 microliters of our mitochondrial DNA. After we ran the gel at 200 volts for 3 minutes. When the gel was complete, it was stained and then we analyzed it the next day.

Results

Discussion
After analyzing our results, it turns out that I do not have the "disease" that we were testing for. Although there were no obvious errors in our lab, one source of error could be pipetting DNA as well as matrix beads into the PCR tube and then having the DNA be destroyed.