Improvements in allelic discrimination of microsatellite markers using denaturing polyacrylamide gel electrophoresis

Poor resolution, retarded progress of DNA through gels, and variable sizing of DNA fragments between and within gels hinder accurate genotyping of som e simple sequence length polymorphism (SSLP) markers with the Perkin Elmer Applied Biosystems 377 Sequenator. These problems are similar to renaturati on related problems observed in DNA sequencing gels. PCR products especiall y susceptible to these problems are shown to have higher melting temperatur es (T-m) than others. Gels containing increased concentrations of denaturan ts allow greater accuracy in allelic discrimination. This is especially ben eficial where quantification is necessary.