Single-tube single-colour multiplex PCR amplification of ten polymorphic microsatellites (ALF10): a new powerful tool for DNA profiling

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Abstract
We studied the genomic diversity of a sample of the Brazilian population with 10 highly polymorphic autosomal microsatellite loci, each located in a different chromosome. The set of 10 microsatellites (ALF10) could be amplified by PCR in a single multiplex reaction and analysed in a single run in a DNA fluorescent automatic sequencer. The discrimination power was very high: the probability of two randomly chosen Brazilian individuals presenting different genetic profiles when typed at the 10 loci was 0.9999999999997. Thus, ALF10 was capable of defining genetic individuality. The expected power of exclusion for paternity testing was 0.99998. In 350 paternity trios in which paternity had been previously ruled out by other DNA tests, exclusions at one or more of the microsatellite loci of ALF10 were observed in all cases (100%). This simple single-tube, single-fluorophore, single electrophoretic run multiplex set of 10 microsatellites constitutes a new powerful tool for DNA profiling, highly useful for paternity testing, forensic studies and human evolutionary studies.

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