Incorporating mathematics principles into high school biology experiments using GFP denaturation studies

Increasing mathematics proficiency among high school students is a priority of our Federal and many state governments. Our goal was to develop a low‐cost experiment in which high school biology teachers with budget constraints could incorporate mathematics principles into their biology experiments in both an interesting and exciting manner. In our experiment, we wanted to determine if we could develop a protocol to measure a statistical difference in fluorescence between folded and fluorescing Green Fluorescent Protein (GFP) and the denatured protein. In order to denature GFP, we used various household chemicals which would be both accessible, cheap, and of which disposal would not be costly. Samples containing lysate obtained from cells expressing GFP in the absence or presence of denaturant were transferred to a microplate and exposed to UV light and photographed using the BlueView Transilluminator. The image was converted to black and white and then analyzed using ImageJ software. Student t tests were performed to determine if it was possible to quantify the difference between the folded and fluorescing and unfolded protein. We determined that statistically significant differences were observed between GFP in water (positive control) and GFP exposed to various denaturants (i.e. heat, vinegar, and others). Importantly, specialized equipment was not required as the transilluminator is both multifunctional and relatively inexpensive. In addition, ImageJ is downloadable free from the NIH and Excel can be used to conduct the t test.