The United States of America as Represented by the Secretary of

The Government of the United States of America, as Represented by the

Mihael H Polymeropoulos

Carl R Merril

Christian Lavedan

Louis William Licamele

Simona Volpi

Changfu Xiao

Christos Polymeropoulos

Vasilios Polymeropoulos

Louis Licamele

The invention relates to polymorphic markers (two tetranucleotide and one dinucleotide repeat polymorphisms) that are useful for human individualization. Applications are in forensic medicine and for paternity and prenatal screening as well as genetic mapping. These markers are characterized by sets of oligonucleotide primers according to the invention useful in PCR amplification and DNA segment resolution. The invention further relates to an assay for measuring the subtle differences in genetic material regarding an added or omitted set of dinucleotide or tetranucleotide repeat polymorphisms which comprises obtaining an amount of nucleotide segments effective for testing, amplifying the segments by the PCR procedure using at least one primer nucleotide sequence according to the present invention, resolving the amplified segments using gel electrophoresis, and comparing the resolved segments by autoradiography to observe the differences in migration patterns due to structural differences. The assay according to the invention is easy to perform and results can be obtained within 24 hours. It is not uncommon for results to be available within 3-4 hours. Accordingly, the invention also relates to an improved PCR procedure and a PCR assay kit which comprise nucleotides according to the invention.

Three highly informative microsatellite repeat polymorphic DNA markers

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Bethesda, MD, United States of America

Mihael H Polymeropoulos