This system

integrates routine laboratory steps by performing cell lysis, DNA isolation, STR amplification, electrophoretic separation, fluorescent detection, and data analysis to generate DNA profiles in under two hours. Previously, PowerPlex® 16 HS chemistry (Promega Corp., Madison, WI), a find more 16 marker assay, was validated on the RapidHIT System [3] and [4]. However, the Federal Bureau of Investigation (FBI), European Network of Forensic Science Institute (ENFSI) and European DNA Profiling Group (EDNAP) have all agreed to the addition of STR loci to the European Standard Set (ESS) and to the core CODIS loci to increase cross-border data sharing, increase discrimination power, and reduce adventitious matches [5], [6], [7] and [8]. Furthermore, the Prüm treaty [9] and [10] was enacted into European 3-Methyladenine in vivo Union legislation which requires member states to submit the five additional loci that are part of the new expanded 12 ESS [11]. This led manufacturers to develop and commercialize products that include all the required and/or recommended loci as requested by ENFSI, EDNAP and the core CODIS Loci Working group [12] and [13]. The GlobalFiler Express PCR Amplification Kit from ThermoFisher Scientific (Waltham, MA),

an NDIS approved chemistry, contains all the required and recommended loci [6] and [8]. The kit contains 21 autosomal STR loci (D3S1358, vWA, D16S539, CSF1PO, TPOX,

D8S1179, D21S11, D18S51, D2S441, D19S433, TH01, FGA, D22S1045, D5S818, D13S317, D7S820, SE33, D10S1248, D1S1656, D12S391, D2S1338) and 3 sex determining markers (Amelogenin, DYS391, Y-indel). Use of fluorescent six-dye technology enables the amplicon sizes to be less than 400 bp (except SE33, 442 bp). To support the community worldwide, increase level of discrimination, facilitate international DNA profile comparison, and reduce risk of adventitious matches, the GlobalFiler Express assay was integrated and validated on the RapidHIT System. The developmental validation experiments presented here were performed according to the quality assurance standards issued by ioxilan the Director of the FBI [14] and the revised guidelines published by the Scientific Working Group on DNA Analysis (SWGDAM) [15]. The results confirm the reliability of the NDIS-approved GlobalFiler® Express assay on the RapidHIT System for generating DNA profiles from reference samples. The profiles can be uploaded after forensic expert review to national and international databases once laboratories have completed their internal validation. Buccal swab samples were collected from consenting donors using 3 inch cotton-tipped swabs from Puritan Medical Products Company (Guilford, ME). Each donor was instructed to swipe the inside of the cheek ten times and contribute swabs daily to generate aged swabs for stability studies.