Appunti di Principi-BiolMol CTF - Capitolo3 - Omero

PCR-RFLPThe polymorphism results from a single nucleotide differencethat provides a recognition site for a restriction enzyme in oneallelic form and not the other. A polymorphism of this type canbe rapidly detected by (1) amplifying the region around thepolymorphic site from each sample, (2) subjecting the amplifiedmaterial to the appropriate restriction enzyme for a brief periodof digestion, and (3) distinguishing the undigested PCR productfrom the smaller digested fragments by gel electrophoresis. Bychoosing primers that are relatively equidistant to andsufficiently far from the polymorphic site, one can easily resolveallelic forms on agarose or polyacrylamide gels.