Application of Molecular Techniques in Forensic Identification: From DNA Profiling to Next-Generation Sequencing

Abstract

Forensic identification has experienced a profound transformation with the advent of molecular biology techniques, shifting from traditional morphological and serological methods to highly sensitive and specific DNA-based approaches. This review provides a comprehensive overview of the evolution of molecular techniques in forensic science, beginning with conventional DNA profiling methods such as Restriction Fragment Length Polymorphism (RFLP) and Short Tandem Repeat (STR) analysis, and extending to advanced methodologies including mitochondrial DNA (mtDNA) analysis, Y-chromosome markers, and Next-Generation Sequencing (NGS).

The review critically examines the principles, applications, advantages, and limitations of these techniques in diverse forensic contexts, including crime scene investigation, human identification, kinship analysis, and mass disaster victim identification. Furthermore, it highlights recent advancements in forensic genomics, epigenetics, and microbiome-based approaches, along with the growing role of bioinformatics and artificial intelligence in data interpretation. Despite significant progress, challenges such as data complexity, ethical considerations, and the need for global standardization remain key concerns. Overall, this review underscores the pivotal role of molecular techniques in modern forensic science and emphasizes their continued evolution toward more precise, rapid, and comprehensive identification systems.