RFLPbased

RFLP-based methods refer to approaches that detect genetic variation by analyzing differences in DNA fragment lengths produced when DNA is cut with restriction enzymes. Polymorphisms may create, destroy, or alter restriction sites, leading to distinct patterns of fragment lengths after enzymatic digestion. The resulting banding patterns can be used to distinguish alleles, individuals, or strains.

Two common implementations exist. In the classic RFLP technique, DNA is digested with restriction enzymes, separated by gel electrophoresis, and the fragments are transferred to a membrane and probed with a labeled DNA sequence that targets a specific locus (Southern blotting). In PCR-RFLP, a region of interest is first amplified by PCR, and the PCR product is then digested with restriction enzymes to reveal allelic variation. In both cases, differences in fragment sizes or the presence/absence of fragments indicate polymorphisms.

RFLP-based methods have played a significant historical role in genetics. They were widely used for genetic mapping, forensics, paternity testing, and microbial or plant strain typing. As technology progressed, PCR-based markers and sequencing approaches—such as AFLP, SSRs, SNP arrays, and next-generation sequencing—offered higher throughput and greater efficiency, reducing reliance on traditional RFLP procedures.

Today, RFLP-based approaches are less common in routine diagnostics and large-scale studies but remain a foundational concept in molecular genetics. They illustrate how variation at restriction sites translates into detectable differences in DNA fragment patterns and provide a bridge between early molecular techniques and modern genotyping methods.