Practice question

Restriction Fragment Length Polymorphism represents one of earliest molecular markers, based on alteration of restriction enzyme recognition sites by mutation, or variation in distance between sites. Genomic DNA is digested with restriction endonuclease, separated by gel electrophoresis, blotted and hybridized with labeled probe following Southern hybridization. Polymorphism appears as fragment length differences. This method does not involve polymerase chain reaction amplification for detection, unlike SSR, RAPD and SNP assays where PCR is integral. Classic RFLP workflow relies purely on digestion and hybridization for genotype visualization.