Optimizing Magnetite Synthesis for DNA Extraction: A Factorial Design Analysis of Temperature, pH, and TEOS Ratio Effects on Yield and Purity

Background: Magnetite nanoparticles are widely used in DNA extraction due to their magnetic properties. Co-precipitation is a commonly used synthesis method, yet the influence of specific synthesis parameters on DNA extraction performance remains unexplored. Methods: This study investigated a factorial design to evaluate the effects of temperature, pH, and tetraethyl orthosilicate (TEOS) ratio on DNA extraction performance, measured by yield and purity. Magnetite was synthesized under various conditions, and DNA was extracted and analyzed for quantity and quality, including PCR amplifiability using a 16S rRNA primer. Results: Among the parameters tested, only the TEOS ratio significantly influenced DNA yield and purity, while temperature and pH showed no significant effects. The optimal conditions identified were 25 °C, pH 10, and a TEOS ratio of 0.25, yielding the highest DNA quality, which could be successfully amplified using PCR. Conclusion: These findings suggest that while co-precipitation is robust, further optimization of the TEOS ratio could enhance DNA extraction efficiency. Future research should explore additional factors influencing magnetite’s DNA extraction capabilities.