HIGH-YIELD OF RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISMS IN 2-DIMENSIONAL SEPARATIONS OF HUMAN GENOMIC DNA

We have investigated the extent to which restriction fragment length p olymorphism can be detected by two-dimensional electrophoresis of end- labeled genomic restriction fragments. Genomic DNA was digested with N otI and EcoRV and labeled at the NotI recognition site before first-di mension electrophoresis in disk gels. DNA in the disk gels was further digested in situ with HinfI prior to second-dimension electrophoresis , yielding patterns in which approximately 2000 end-labeled fragments were simultaneously visualized. On the basis of studies of 6 mother/fa ther/child trios, a group of 184 fragments was organized into 85 polym orphic systems in which all allelic fragments were detectable in the 2 -D patterns. Another 206 fragments varied as to their presence among i ndividuals, but their relatedness to other fragments was not establish ed. Our data indicate that a large number of DNA polymorphisms can be simultaneously scored in 2-D separations of genomic DNA fragments. (C) 1995 Academic Press, Inc.