Abstract

The purpose of our research was to design primers and experimentally identify PCR conditions that will allow us to view bands of the mutation A455E on an agarose gel through PCR analysis. Cystic Fibrosis is a chronic disease that greatly affects those of Caucasian descent (Braekeleer et al. 1997). The mutation A455E results in a milder form of CF than other mutations (Braekeleer et al. 1997).  Designing primers that specifically bind to the mutation if it is present in the DNA can help determine whether a patient has a specific mutation. The forward and reverse primers were created using the CF mutation database. They will bind to the mutation and a band will be present in the gel if the tested human bronchial cells contained the A455E mutation. The control primer will bind to the DNA if no mutation is present. We hypothesized that by having the mutation at the end of the forward primer, the Taq polymerase is more likely to recognize and bind to the primer and the DNA sequence than if the mutation was in the middle of the primer. The presence of the band at 831 base pairs showed that the mutation was present in the S9 cells tested. For the DIY lab, a survey was created to compare Cystic Fibrosis to AIDS, cancer, heart disease, and diabetes. Lyman Briggs students were surveyed about the diseases. Analysis of the survey showed that cancer and heart disease are considered to be two of the most detrimental diseases.