Bacterial resistance to antibiotics is an escalating global health concern, primarily driven by genetic mutations that enhance survival against antimicrobial regimes. Understanding the mechanisms of antibiotic resistance is vital for developing effective strategies to combat antimicrobial resistance (AMR). Several bioinformatics tools and databases, such as AMRFinder, ResFinder, and the Comprehensive Antibiotic Resistance Database (CARD), play a significant role in identifying resistance genes from whole-genome sequencing data. CARD offers a robust platform with features including resistome profiling, resistance gene identification, and visualization tools, facilitating an in-depth analysis of AMR mechanisms. This chapter presents a step-by-step methodology for using CARD to identify AMR genes, providing a clear framework for researchers to analyze resistance determinants.