Restriction fragment length polymorphism (rflp)
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Restriction Fragment Length Polymorphism (RFLP) was an early forensic technique using differences in DNA sequences between individuals. RFLP involves digesting DNA with restriction enzymes, separating the fragments by size via electrophoresis, and detecting variants using radioactive probes. Differences in the patterns of fragment sizes between individuals allows for individual identification from biological samples.
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Restriction fragment length polymorphism (rflp)
- 1. Restriction Fragment Length Polymorphism (RFLP)
- 2. Restriction Fragment Length Polymorphism or RFLP was one of the first methods used in forensic investigations, preceding PCR. It is based on the differences in DNA sequences on homologous chromosomes that result in different patterns of restriction fragment length (DNA segments) after restriction enzyme digestion in DNA molecules of different individuals.
- 3. In RFLP, the extracted DNA is digested with specific restriction enzymes. These enzymes recognise and cut the DNA at precise points to produce a collection of DNA fragment of precisely defined lengths. The millions of fragments produced can then be separated by electrophoresis on agarose gels with the smaller fragments migrating farther than the larger fragments.
- 5. One or more of these RFLP fragments can be detected and analysed using nucleic acid probes that are radioactively labelled.
- 6. The RFLP method takes advantage of the polymorphisms present in the genetic code of individuals. The pattern of fragment sizes will differ for each individuals tested. The main advantage of RFLP analysis over PCR- based protocols is that no prior sequence information, nor oligonucleotide synthesis, is required.
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