The use of dinucleotide microsatellites in high-throughput, semi-automated genome mapping

Microsatellites are tandem repeats of simple sequences that occur frequently and at random throughout the human genome. These highly polymorphic markers are flanked by unique sequences which makes them amenable to PCR amplification. Since the microsatellite polymorphism is stable and is inherited in a Mendelian fashion these sequences can be used as highly informative genetic markers. PCR primers can be labeled with one of the four currently available fluorescent dyes. After PCR the products are separated on acrylamide gels. Using a scanning laser and commercially available software primers labeled with different dyes alleles can be distinguished even when their sizes overlap. Allele sizes are reproducibly and accurately determined. We have developed an efficient process, employing currently available technology, to determine accurately large numbers of human genotypes. We are now able to define thousands of individual genotypes per technician per day. This information can be used in genetic studies to define linkage to common human diseases.