Comparative analysis of commercially available metagenomic kits for optimal DNA extraction from bovine fecal samples

Abstract

In the field of metagenomic research, the choice of DNA extraction methods plays a pivotal yet often underestimated role in shaping the reliability and interpretability of microbial community data. This study delves into the impact of five commercially available metagenomic DNA extraction kits on the analysis of bovine fecal microbiota. Recognizing the centrality of accurate DNA extraction in elucidating microbial community dynamics, we systematically assessed DNA yield, quality, and microbial composition across these kits. Notably, the FastDNA spin soil kit yielded the highest DNA concentration, while significant variations in quality were observed across kits. Furthermore, differential abundance analysis revealed kit-specific biases, impacting taxa representation. Microbial richness and diversity were significantly influenced by the choice of extraction kit, with QIAamp DNA stool minikit, QIAamp Power Pro, and DNeasy PowerSoil outperforming the Stool DNA Kit. Principal-coordinate analysis revealed distinct clustering based on DNA isolation procedures, particularly highlighting the unique microbial community composition derived from the Stool DNA Kit. Differential abundance analysis showcased kit-specific biases, influencing taxa representation. This study also addressed practical implications, demonstrating how kit selection influences the accurate identification of spiked bacteria. In essence, our research highlights the need for meticulous consideration of DNA extraction kits in metagenomic studies, offering valuable insights for researchers striving to advance the precision and depth of microbiota analyses in animals.