Isolation of nucleic acids—DNA, RNA, and plasmids is a core technique in plant molecular biology, enabling detailed analysis of genetic information and cellular processes. Plant systems present unique challenges due to rigid cell walls and the presence of interfering compounds such as polysaccharides and phenolics. Effective isolation therefore relies on optimized protocols involving mechanical disruption, chemical lysis, purification, and stabilization steps. Common approaches include CTAB- and SDS-based methods for DNA extraction, guanidine thiocyanate protocols for RNA isolation, and alkaline lysis for plasmid recovery. Careful selection of plant material, particularly young tissues, improves yield and quality. Removal of contaminants using organic solvents and enzymatic treatments such as RNase and DNase ensures purity of the final preparation. Quality assessment using spectrophotometry and gel electrophoresis is essential before downstream applications. Maintaining sterility and preventing nuclease contamination are critical, especially for RNA work. These techniques support a wide range of applications, including PCR, sequencing, gene expression analysis, and genetic engineering, making nucleic acid isolation a foundational step in modern plant biotechnology research.