Testing Diseased Genes Through PCR

Introduction:
Genes, the coding sections of our DNA, are the core of our development. Everything we are, that we become, is in essence controlled by our genes. By studying these genes we unlock the ability to look at life at a developmental stage. Genes control who we are, and they are the key to understanding our diseases. Diseases can be caused by a plethora of reasons, including the faulty production of protein caused by a gene. By looking at a gene from a healthy sample and comparing it to an unhealthy sample we can take a look at the changes in our genetic code that result in these crippling issues. To look at these genes we are using primers to find an intron, PV92, in our genetic code which we will then amplify and study. The "gene" has two possible modes and 3 total combinations. It can either be long or short, and the combinations are homozygous dominant (long), homozygous recessive (short) or Heterozygous.

Purpose:
We do this lab in order to better understand the biological processes that occur in us, and to harness the ability to temper them to our benefit. If we can study these genes and observe the consequences of their malfunction we can gain enormous insight into these disorders. While the lab itself test a noncoding section of our DNA it is easy to extrapolate as to the purpose in testing for genes. We can study genetic predisposition, for parents with a disease, we can look at an individuals risk factors and the necessary procedures to limit them, and a host of other possibilities.

Method:

We start our lab by extracting our DNA sample. By swabbing the inside of our cheeks in order to procure a sample. Following this we will add a protein called Instagene Matrix. The Instangene Matrix beads allow for chelex to protect the DNA from the DNAse that is present in the cell's cytoplasm. We will be putting the sample DNA that we have extracted from the previous step into the PCR. PCR will replicate several times creating a larger sample. This sample will then be placed through Gel Electrophoresis in order to study the "gene" in question. Gel electrophoresis will seperate the sample by size; smaller samples will move further, whereas larger sample will move less.




Discussion:
Our lab results were a catastrophic failure, because of our problems loading the gel, prior to electrophoresis. The well in one of our samples was punctured and the sample rapidly bled through resulting in a complete obscuration of any comparable data between the actual samples of our table. However the gel itself was run correctly, as the controls and the marker were both clearly present. Additionally the sample that we used were extremely faded possessing nowhere near the depth to be clearly seen on the stained gel. There are a few sources of error that could account for our misgivings in this lab. It is possible that we did not extract a proper sample of cheek cells, and even though it was amplified the DNA available was simply not enough. It is possible that the DNA degenerated because we did not properly extract a sample that was only from the supernatent, resulting in DNAse digesting the sample. It is even possible that due to a punctured well there was not enough DNA in the well to properly analyze, or that the bleeding from the bottom of the well resulted in a obfuscation of the results.