Hales, Dale B.
Cell transfection is an exemplary tool in protein analysis and examining transcription factor activity. While general procedures are commonly followed, each cell line requires slight differentiation in pBABE/PEI JET ratios and initial cell count for optimal transfection results. This research focusses on determining a successful procedure for TOV-112 carcinoma cells for future work with 2-methoxyestradiol and NFkB. 2-methoxyestradiol is an angiogenesis inhibitor believed to decrease inflammatory processes. NFkB promoter controls the luciferase gene on the MetLuc plasmid and is pro-inflammatory. The objective of this experiment is to modify the variables until successful results at 24 and 48 hours are observed. Four experimental transfections were conducted during the three months of research, which analyzed the effectiveness of high and low pBABE/PEI JET ratios and increased vs. decreased initial plate seeding. Results show that optimal TOV-112 transfection occurs with a 1:2 pBABE/PEI JET ratio and an initial cell count of around 3x10^5 cells/mg. Future work will hope to emphasize this conclusion and continue with the overarching NFkB ovarian cancer research, which aims to reduce the cases of lethal occurrences.