The Identifiler kit addresses a significant impediment to more rapid genotype processing by eliminating the need for two amplifications and two lanes or injections of electrophoretic analysis to obtain complete CODIS or international profiles for a particular sample. Processing is further enhanced by increased allelic representation in the allelic ladder, decreasing the number of "off-ladder" alleles that must be rerun. In addition to reducing processing steps, the kit has been designed to preserve existing AmpFLSTR kit primer sequences and reaction conditions. In order to verify that the Identifiler kit performs as reliably and robustly as other STR multiplexes, a developmental validation was conducted according to relevant Quality Assurance Standards for Forensic DNA Testing Laboratories. A description of materials and methods addresses locus selection and characterization; PCR amplification; electrophoresis, detection, and analysis; balance calculations; primer set concentration; PCR reaction mix; thermal cycling parameters; species specificity, sensitivity, stability, and mixture studies; and population and concordance studies. The kit was found to meet the manufacturer's claims and is therefore reliable for use in human identity and parentage testing. 7 figures, 3 tables, and 33 references