Browsing by Subject "Biomedical Devices and Instrumentation"
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Item Concordance Study of Forensic Casework Samples Using the AMPFlSTR Kit, AMPFlSTR Identifiler Kit, AMPFlSTR Profiler Plus Kit and PowerPlex 16 Kit(2002-07-01) Armstrong, Treva L.; Arthur Eisenberg; John Planz; Joseph WarrenThe PowerPlex 16, AmpFlSTR Profiler Plus and AmpFlSTR COfiler Kits allow for the co-amplification of the amelogenin gender determining marker and the thirteen core CODIS STR loci: D3S1358, FGA, vWA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, THO1, TPOX and CSF1PO. PowerPlex 16 adds the Penta D and Penta E loci and the AmpFlSTR Identifiler addes the D2S1338 and D19S433 loci. Manufactures of these systems have a suggested input DNA sample range of 0.5-2.5 ng, but have been successfully used to type samples containing less than 0.5 ng of DNA. In this study, several questions were addressed: First, “Are all four STR multiple kits concordant in their reproducibility, reliability, sensitivity and efficiency?” Second, “Is one particular STR megaplex kit more applicable to routine forensic casework?” and Third, “In a mixed DNA sample can individuals, whether male or female, be differentiated?” This paper describes a casework concordance study using adjudicated nonprobative sexual assault, mixed DNA and reference blood samples. Amplifications on all samples, were performed using the AmpFlSTR COfiler, Profiler Plus and Identifiler and PowerPlex 16 Kits and genotyping results were obtained using GeneScan and Genotyper software.Item Evaluation of SPEX6750 Freezer Mill Pulveration of Teeth for Use in Nuclear Casework at the Armed Forces DNA Identification Laboratory(2008-08-01) Bintrim, Kristin A.; John Planz; Arthur Eisenberg; Joseph WarrenBintrim, Kristin A., Validation of Freezer Mill Pulveration of Teeth for use in Nuclear Casework at the Armed Forces DNA Identification Laboratory. Master of Science (Forensic Genetics), August, 2008, 44 pp., 4 tables, 6 figures, bibliography, 27 titles. I completed a validation study at the Armed Forces DNA Identification Laboratory. It was an evaluation of freezer mill pulveration of teeth for use in nuclear casework. The teeth were cleaned using a bleach and sonication method and then ground using a freezer mill. DNA extraction was achieved by using silica based capture techniques. The DNA was quantified, amplified and evaluated using fragment analysis. Several different amounts of input tooth powder were attempted to increase the DNA yield. Qiagen MinElute produced higher DNA yields then Promega’s DNA IQ system for the small sample size tested. After reviewing the results it was concluded that the DNA was most likely highly degraded and therefore did not produce complete genetic profiles for the samples tested.Item Evaluation of the AluQuant Human DNA Quantitation System Using the 96-Well Plate Format(2002-08-01) Alexander, Uvonna Faye Lewallen; John Planz; Arthur EisenbergAlexander, Uvonna Faye Lewallen, Evaluation of the AluQuant Human DNA Quantitation System Using the 96-Well Plate Format. Master of Science (Forensic Genetics), August 2002, 121 pp., 4 charts, 4 tables, 8 figures, 2 appendices, references, 13 titles. This study evaluated the AluQuant Human DNA Quantitation System (Promega Corporation, Madison, WI) using the 96-well plate format for possible implementation by Orchid Cellmark Dallas. The importance of human DNA quantitation in forensics is two-fold. First, the Quality Assurance Standards set forth by the DNA Advisory Board requires human DNA in forensic samples be quantitated. Also, the highly sensitive PCR multiplex PCR multiplex assays used in forensics have been optimized for a narrow range of template DNA, thus requiring accurate and consistent quantitation. This evaluation consisted of three general goals: examination of the Reporter Microplate Luminometer (Turner BioSystems, Sunnyvale, CA), alteration of the assay variables to obtain optimal performance, and characterization of the assay. The Reporter produces reproducible results and is sensitive to at least 4.88 x 10^-9 moles ATP. Of the variables tested, quick centrifugation of the incubation plate had the most noticeable effect on the results obtained. The assay did not perform as characterized by Promega. AluQuant is not reproducible, nor does it consistently produce results within a two-fold accuracy range. Therefore, Orchid Cellmark Dallas will not be implementing the AluQuant Assay.Item Synergy 2007: Annual Research Report(2007-01-01)Item The Evaluation of the Maxwell 16 and the DNA IQ Casework Sample Kit for the Extraction of DNA from Forensic Samples(2007-08-01) Alkhazin, Tarig; Arthur Eisenberg; Joseph Warren; John PlanzThe Maxwell 16 (Promega Corporation, Madison, WI) is a small, self-contained instrument utilizing Promega’s DNA IQ chemistry for the automated extraction of DNA from 16 biological samples simultaneously. Currently, the Maxwell 16 is used in conjunction with the DNA IQ Reference Sample kit for the automated extraction of DNA from forensic and paternity reference samples. Promega Corporation is currently in the development of the DNA IQ Casework Sample kit with the intent of using the Maxwell 16 instrument in the extraction of DNA from forensic evidentiary samples. Modifications have been made to Maxwell 16 to allow the elution of DNA in a smaller volume (Low-Elution Volume (LEV) configuration) that when used in conjunction with the DNA IQ Casework Sample kit would optimize DNA yield from forensic casework samples. However, the limited quantity and the low quality of forensic casework samples are significant challenges facing most automated DNA extraction systems. An evaluation study was conducted to test the performance of the Maxwell 16 instrument along with the DNA IQ Casework Sample kit for processing forensic evidentiary samples. Mock evidentiary items used for the evaluation consisted of blood, semen, tissue, and touch samples that are routinely encountered in forensic casework. Prior to loading the Maxwell 16 instrument samples were first digested using the Tissue and Hair Extraction kit (Promega Corporation) designed for optimum DNA recovery. The extraction of DNA from sexual assault samples typically requires the separation of the sperm DNA deposited by the assailant from the vaginal epithelial cell DNA from the victim. Promega Corporation has developed the Differex System which utilizes a manual phase separation technique to obtain both a sperm fraction and an epithelial cell fraction. After separation and digestion, samples were then added to the DNA IQ Casework Sample kit cartridges for automated DNA purification using the Maxwell 16. For comparison, DNA was also obtained from replicate samples processed with the Differex System following the standard organic extraction method. The evaluation of the Differex system with mock sexual assault samples was conducted in order to determine if this differential extraction process can be used in conjunction with Maxwell 16 to improve case processing efficiency. To evaluate the performance of the Maxwell 16 and the DNA IQ Casework Sample kit, replicate samples were prepared and processed using UNTHSC’s standard organic extraction methodology. The DNA obtained by both methodologies was quantified using the Applied Biosystems (AB) Quantifiler Human DNA Quantification kit (Foster City, CA) and the AB 7500 Real-Time PCR system and then amplified using the PowerPlex 16 kit (Promega Corporation). The amplified DNA was analyzed using the AB 3130xl Genetic Analyzer and the resulting STR electropherograms were analyzed to obtain profiles using GeneMapper ID v3.2 (Applied Biosystems). In addition to assessing the quantity and the quality of the DNA obtained, blank cartridges were simultaneous processed with the mock forensic samples to demonstrate whether the Maxwell 16 could introduce cross-contamination between samples. The overall performance and the cost-effectiveness are the ultimate criteria to help determine the utility of the Maxwell 16 in the processing of forensic evidentiary casework samples.