title: Developmental validation of the PowerPlex(®) 21 System.
abstract: The PowerPlex(®) 21 System is a STR multiplex that has been optimized for casework samples while still being capable of database workflows including direct amplification. The loci included in the multiplex offer increasing overlap with core loci used in different countries and regions throughout the world. The PowerPlex(®) 21 System contains D1S1656, D2S1338, D3S1358, D5S818, D6S1043, D7S820, D8S1179, D12S391, D13S317, D16S539, D18S51, D19S433, D21S11, Amelogenin, CSF1PO, FGA, Penta D, Penta E, TH01, TPOX, and vWA. These loci represent all 13 core CODIS loci in addition to loci commonly used in Asia and Europe. A developmental validation study was completed to document performance capabilities and limitations of the PowerPlex(®) 21 System. Data from this validation work served as the basis for the following conclusions: genotyping of single-source samples was reliable across a range of template DNA concentrations with >95% alleles called at 50 pg. Direct amplification of samples from FTA(®) storage cards was successfully performed using the reagents provided with the system and modified cycling protocols provided in the technical manual. Mixture analysis showed that over 95% of minor alleles were detected at 1:9 ratios. Reaction conditions including volume and annealing temperature as well as the concentrations of primers, DNA polymerase, magnesium, and Master Mix were shown to be optimal and able to withstand moderate variations without affecting system performance. Reproducible results were generated by different users at different sites. Finally, concordance studies showed consistent results when comparing the PowerPlex(®) 21 System with other commercially available STR-genotyping systems.
authors: Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, United States. Electronic address: martin.ensenberger@promega.com.
authors: Martin G
authors: MG
authors: Ensenberger
authors: National Institute of Standards and Technology, 100 Bureau Drive, M/S 8312, Gaithersburg, MD 20899, United States.
authors: Carolyn R
authors: CR
authors: Hill
authors: Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, United States.
authors: Robert S
authors: RS
authors: McLaren
authors: Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, United States.
authors: Cynthia J
authors: CJ
authors: Sprecher
authors: Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, United States.
authors: Douglas R
authors: DR
authors: Storts
mesh_terms: D000818:Animals; D002176:Candida albicans / Q000235:genetics; D002415:Cats / Q000235:genetics; D002417:Cattle / Q000235:genetics; D002645:Chickens / Q000235:genetics; D004247:DNA / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods*; D003670:Deer / Q000235:genetics; D004285:Dogs / Q000235:genetics; D019075:Electrophoresis, Capillary; D005456:Fluorescent Dyes; D005819:Genetic Markers; D006090:Gram-Negative Bacteria / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000235:genetics; D006736:Horses / Q000235:genetics; D006801:Humans; D051379:Mice / Q000235:genetics; D018895:Microsatellite Repeats; D060885:Multiplex Polymerase Chain Reaction / Q000295:instrumentation*; D011817:Rabbits / Q000235:genetics; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013045:Species Specificity; D013552:Swine / Q000235:genetics
chemical_list: D005456:Fluorescent Dyes; D005819:Genetic Markers; D004247:DNA
keywords: CODIS; DNA typing; Forensic science; PowerPlex(®); Short tandem repeat (STR); Validation