Variations in data obtained from experiments often correlate with the lack of standardized protocols for equal measurement. The iGEM Interlab Study serves the purpose of tackling this problem by providing a standard measurement protocol for all teams to follow in a controlled setting, in order to effectively characterize the increased efficiency of doing the same. Green Fluorescent Protein (GFP) is a biomarker that is commonly used in labs across the world, and for this purpose, it was used as the standard gene of expression due to the shared capability of most teams to quantify it. Through this standardized protocol, each team appropriately quantified GFP from their transformations and liquid cultures, and the data from each team serves as levels of treatment to compare and improve the measurement and lab protocols used. Our team collaborated with the University of Georgia for access to a plate reader and for a comparison of measurements. Following iGEM’s transformations protocol, we transformed the samples at Lambert High School and transported the plates to the University of Georgia for the inoculation of liquid cultures and measurement. Safety precautions were taken in each step to ensure prevention of contamination and proper enclosement of bacterial samples.
The LUDOX Solution provided to our team was stored in a freezer for an extended period of time, eventually rendering in dysfunctional. The fluorescein solution also appeared to not express color as expected. Due to this, we utilized the LUDOX solution and fluorescein provided to the UGA team in order to obtain our OD Reference and fluorescein standard curves. The transformations were conducted at Lambert High School, plated and parafilmed, and transported to UGA for inoculation. During sample measurement, we obtained several OVRFLW errors in certain samples, and despite changing gain settings, the error was still registered. We were unable to obtain data for those replicates but did obtain data for the others. We utilized UGA’s Biotek Synergy H4 Plate Reader for our measurements and conducted all measurements under the supervision of the UGA iGEM Team and their advisors.
Refer below for the data we collected. (There is more than one page.)