Several materials were evaluated to demonstrate genotyping reproducibility and reliability. Five sites evaluated panels of extracted VX-809 DNA, buccal Indicating FTA® cards, buccal cotton swabs, and nonFTA Bode Buccal DNA Collectors™ with three replicates for each sample. Samples were detected using 3130 and 3500 Series Genetic Analyzers or a 3730 DNA Analyzer.
Five sites evaluated the NIST SRM2391c PCR-Based DNA Profiling Standard samples A–D. Complete and concordant profiles were gathered at each of the five test sites for all samples (n = 72), except with sample D. Sample D was a mixture sample with four alleles at D12S391: 18.3, 19, 22, and 23. All alleles were consistently called except the 19 allele. Although the 19 allele resolved as a distinct shoulder on the 18.3 allele peak, neither the GeneMapper®ID nor the GeneMapper®ID-X software called the minor contributor 19 allele ( Fig. 3). Similar resolution was seen across all replicates on the 3130 and 3500 Series Genetic Analyzers and a 3730 DNA Analyzer, and can be expected with closely spaced minor contributor alleles. Complete and concordant profiles were gathered from multiple solid support substrates. All
five buccal cotton swab samples gave full and concordant profiles from both test sites (n = 45). A complete and concordant profile was seen for four buccal Indicating FTA® card samples and SRM2391c sample F (cells spotted onto an FTA® card) at each of four test sites (n = 70). Five nonFTA punches from four Bode Buccal DNA Collectors™ and the SRM2391c sample E (cells spotted onto S&S 903 paper) Alisertib solubility dmso gave full and concordant profiles (n = 54). Two of the sample sources, one FTA® card and one Bode Buccal DNA Collector™, produced low peak heights at each evaluation site, presumably due to poor cell transfer onto the surface or low shedding of buccal cells from the donor. Any partial profile samples were fully concordant at all amplified loci. Artifacts specific to the migration of PowerPlex® Fusion System amplification products on POP-7™ polymer were observed. Artifacts
were labeled by the GeneMapper®ID Software, version 3.2, at approximately 88 bases in the fluorescein channel and approximately 90 bases in the JOE channel. All samples except allelic ladder contained the artifacts, including negative controls. Artifacts crotamiton may be reduced by performing sample electrophoresis immediately after amplification. These artifacts were not observed on POP-4® polymer and are noted in the technical manual [9]. Forensic casework samples represent a wide variety of sample quantity, background contaminants, and biological sample types. Four validation sites evaluated a total of 76 case-type samples from their own collections (Table 1). Samples were extracted from a variety of sources by organic and EZ1® extraction methods. Detection was performed on either an Applied Biosystems® 3130 or 3500 Series Genetic Analyzer, and data was analyzed with GeneMapper® ID-X software.