Single-tube single-colour multiplex PCR amplification of 10 polymorphic microsatellites (ALF10): a new powerful tool for DNA profiling

We studied the genomic diversity of a sample of the Brazilian population wi th 10 highly polymorphic autosomal microsatellite loci, each located in a d ifferent chromosome. The set of 10 microsatellites (ALF10) could be amplifi ed 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 d ifferent genetic profiles when typed at the 10 loci was 0.9999999999997. Th us, 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, exclusion s at one or more of the microsatellite loci of ALF10 were observed in all c ases (100%). This simple single-tube, single-fluorophore, single electropho retic run multiplex set of 10 microsatellites constitutes a new powerful to ol for DNA profiling, highly useful for paternity testing, forensic studies and human evolutionary studies.