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Next-generation sequencing (NGS) has transformed genomics, presenting significant potential for advancing wildlife conservation. NGS technology offers opportunities and challenges for addressing scientific questions in wildlife management. Unfortunately, there is a lack of comprehensive records on NGS implementation in Southeast Asia, particularly concerning wildlife conservation. To address this gap, we analyzed NGS studies focused on wildlife monitoring in Southeast Asia and introduced a practical framework for implementing NGS technologies in global wildlife conservation, especially in Southeast Asia. We systematically reviewed NGS studies in wildlife monitoring from the SCOPUS database. We identified 137 relevant publications from 11 countries, with Malaysia contributing 36% of the studies. The included studies were categorized into five themes: species identification, dietary assessment, health …