Microbial DNA quantification using spectrophotometric and fluorometric methods using magnetic nanoparticle-based extraction

This study evaluates the precision of three DNA quantification methods—spectrophotometry, fluorometry, and qPCR using Cycle Threshold (Ct)—using a magnetic nanoparticle-based extraction kit from KODC on microbial DNA. Our results indicate that the qPCR method exhibits the highest precision, characterized by the lowest coefficient of variation (CV) and standard deviation (SD), making it the most consistent choice for routine laboratory applications. In contrast, fluorometry shows moderate precision, while spectrophotometry demonstrates the least reliability due to higher variability. Mathematical modeling further revealed both spectrophotometry and fluorometry exhibited weak correlations with each other and with qPCR, limiting their predictive capabilities. Notably, the Generalized Additive Model (GAM) provided the best fit for the data, highlighting its effectiveness in capturing underlying trends. These findings underscore the importance of method selection based on precision needs and suggest that standardizing protocols could enhance consistency in DNA quantification. Further investigations are warranted to explore the impact of varying conditions on these methods and to refine the understanding of their relationships.