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RFLP (Restriction Fragment Length Polymorphism ) Introduction RFLP is one of the earliest DNA-based molecular marker techniques. It detects variation in DNA sequences between individuals, caused by mutations, insertions, deletions, or restriction enzyme recognition site changes. First developed in late 1970s–1980s. Used extensively in genetic mapping, DNA fingerprinting, and gene detection. It is co-dominant, locus-specific, and highly reproducible. Principle RFLP is based on the fact that DNA from different individuals may differ in the presence or absence of restriction enzyme recognition sites, leading to different fragment sizes after digestion. Key idea: DNA → Restriction enzyme digestion → Fragments → Gel separation → Detection using labeled probe → Polymorphism analysis Materials Required Genomic DNA from the organism Restriction enzymes (e.g., EcoRI, HindIII) Agarose or polyacrylamide gel Nylon or nitrocellulose membrane Radioactive or non-radioactive DNA probe Autoradiography ...