Temporal dynamics of rat auricle skin microbiota during early decomposition and association with postmortem interval estimation v2

<p>Thisprotocol describes a longitudinal study of microbial succession on rat auricleskin during early postmortem decomposition (1 h to 5 d). The workflow includesanimal euthanasia, repeated skin swabbing at four postmortem intervals (1 h, 1d, 3 d, 5 d), DNA extraction, 16S rRNA gene sequencing (V3–V4 region), andbioinformatic analysis using QIIME2 and DADA2. The resulting taxonomicabundance tables are used for machine learning-based postmortem interval (PMI)prediction using Ridge regression and Random Forest, with leave-one-subject-outcross-validation (LOSO-CV) to prevent data leakage [1].</p>