What is restriction fragment analysis

And that results in a polymorphism, or difference between those two people. We typically see these, or we monitor these, by isolating the DNA, cutting it with that bacterial restriction enzyme, and running it on a gel using electrophoresis. In one person, without the enzyme site you'll see one band, and the person that has the enzyme site, you'll see two bands, representing the two cleaved products.

So these differences in nucleic acid sequences and restriction enzyme binding sites just mean that there's a difference in the sequence between those two people. That sequence difference doesn't necessarily mean that there's a disease associated with it. It's a polymorphism that we could use to follow the inheritance of DNA. The digested DNA is size-fractionated on a preparative agarose gel, and fragments ranging from to bp are excised, eluted and cloned into a plasmid vector for example, pUC Digests of the plasmids are screened to check for inserts.

Southern blots of the inserts can be probed with total sheared DNA to select clones that hybridize to single- and low-copy sequences. You are here: NCBI. External link.

Larger quantities are needed for species with larger genomes, and for the greater number of times needed to probe each blot. RFLPs are not amenable to automation and collaboration among research teams requires distribution of probes. Applications RFLPs can be applied in diversity and phylogenetic studies ranging from individuals within populations or species, to closely related species.

RFLPs have been widely used in gene mapping studies because of their high genomic abundance due to the ample availability of different restriction enzymes and random distribution throughout the genome. They also have been used to investigate relationships of closely related taxa, as fingerprinting tools, for diversity studies, and for studies of hybridization and introgression, including studies of gene flow between crops and weeds.

Suggested reading Restriction fragment length polymorphism mapping in conifers and applications to forest genetics and tree improvement Neale, D. Twitter Whatsapp Linkedin Email.