Methods matter: Assessing the efficiency of extraction and library preparation protocols in archaeological dental calculus analysis

Recent developments in DNA extraction and library preparation techniques, in conjunction with advancements in next generation sequencing and bioinformatic tools, have broadened the horizons of research aimed at exploring the correlation between the oral microbiome and ancient health, dietary habits, and lifestyles. However, various protocols within the DNA extraction and library preparation process can influence the effectiveness of recovering the microbial composition derived from archaeological dental calculus samples. To the best of our knowledge, there is a dearth of research that has systematically evaluated the combined performance of these protocols on samples derived from various preservation states. This critical gap in the existing literature underscores the need for this study. Here, we compare commonly used extraction and library preparation methods on dental calculus samples of varying preservation states.