title: Developmental validation of the PowerPlex(®) ESI 16/17 Fast and PowerPlex(®) ESX 16/17 Fast Systems.
abstract: The PowerPlex(®) ESI 16 Fast, ESI 17 Fast, ESX 16 Fast, and ESX 17 Fast Systems represent faster cycling versions (50min or less) of the PowerPlex(®) ESI and ESX Systems released by Promega in 2009 to accommodate the ENFSI and EDNAP groups' call for new STR multiplexes for Europe. In addition to amplification of purified DNA samples, these new faster cycling systems allow for direct amplification from single-source blood and buccal samples deposited on FTA(®) and nonFTA paper as well as from SwabSolution™ extracts of buccal swabs without the need for purification and quantitation. There are no changes to the autosomal primer pair sequences in the PowerPlex(®) ESI Fast and ESX Fast Systems compared to the original multiplexes, and full concordance at all autosomal loci and amelogenin was observed with data generated previously with the original PowerPlex(®) ESI and ESX Systems. This paper describes the developmental validation study performed on these new fast systems following guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM) and those of the DNA Advisory Board (DAB). Validation data demonstrate that these systems are sensitive for detecting low levels of DNA while also being capable of generating robust profiles from the high amount of input DNA present in direct-amplification samples. These systems are also tolerant to both high concentrations of PCR inhibitors as well as to slight variations in the final concentration of master mix and primer pair present in the amplification reaction that might be encountered due to pipetting error. The results of this validation study demonstrate that these systems may be used on multiple thermal cyclers and capillary electrophoresis platforms.
authors.affiliation: Promega Corporation, Madison, WI 53711-5399, USA. Electronic address: bob.mclaren@promega.com.
authors.forename: Robert S
authors.initials: RS
authors.lastname: McLaren
authors.affiliation: Promega Corporation, Madison, WI 53711-5399, USA.
authors.forename: Jeanne
authors.initials: J
authors.lastname: Bourdeau-Heller
authors.affiliation: Promega Corporation, Madison, WI 53711-5399, USA.
authors.forename: Jaynish
authors.initials: J
authors.lastname: Patel
authors.affiliation: Promega Corporation, Madison, WI 53711-5399, USA.
authors.forename: Jonelle M
authors.initials: JM
authors.lastname: Thompson
authors.affiliation: Key Forensic Sciences, 207 B/C Cavendish Place, Birchwood Park, Warrington, Cheshire, WA3 6WU, UK.
authors.forename: Jenny
authors.initials: J
authors.lastname: Pagram
authors.affiliation: Key Forensic Sciences, 207 B/C Cavendish Place, Birchwood Park, Warrington, Cheshire, WA3 6WU, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Loake
authors.affiliation: Key Forensic Sciences, 207 B/C Cavendish Place, Birchwood Park, Warrington, Cheshire, WA3 6WU, UK.
authors.forename: David
authors.initials: D
authors.lastname: Beesley
authors.affiliation: National Bureau of Investigation, Forensic Laboratory (NBI), Jokiniemenkuja 4, 01301 Vantaa, Finland.
authors.forename: Markus
authors.initials: M
authors.lastname: Pirttimaa
authors.affiliation: Biomolecular Measurement Division, National Institutes of Standards and Technology (NIST), 100 Bureau Drive, Gaithersburg, MD 20899-8312, USA.
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Hill
authors.affiliation: Biomolecular Measurement Division, National Institutes of Standards and Technology (NIST), 100 Bureau Drive, Gaithersburg, MD 20899-8312, USA.
authors.forename: David L
authors.initials: DL
authors.lastname: Duewer
authors.affiliation: Biomolecular Measurement Division, National Institutes of Standards and Technology (NIST), 100 Bureau Drive, Gaithersburg, MD 20899-8312, USA.
authors.forename: Margaret C
authors.initials: MC
authors.lastname: Kline
authors.affiliation: Biomolecular Measurement Division, National Institutes of Standards and Technology (NIST), 100 Bureau Drive, Gaithersburg, MD 20899-8312, USA.
authors.forename: John M
authors.initials: JM
authors.lastname: Butler
authors.affiliation: Promega Corporation, Madison, WI 53711-5399, USA.
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Storts
mesh_terms: D000818:Animals; D053836:DNA Degradation, Necrotic; D016172:DNA Fingerprinting; D019075:Electrophoresis, Capillary; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D060885:Multiplex Polymerase Chain Reaction / Q000295:instrumentation*; D015203:Reproducibility of Results; D013045:Species Specificity; D013048:Specimen Handling / Q000379:methods
keywords: Direct; ESI; ESX; Fast; PowerPlex(®); Validation