Browsing by Subject "Validation"
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Item The Validation of Applied Biosystems Quantifiler Human DNA Quantification Kit and Quantifiler Y Human Male DNA Quantification Kit for the Armed Forces DNA Identification Laboratory(2004-08-01) Levandowsky, Elizabeth C.; Arthur Eisenberg; Joseph Warren; John PlanzLevandowsky, Elizabeth C., The Validation of Applied Biosystems Quantifiler™ Human DNA Quantification Kit and Quantifier™ Y Human Male DNA Identification Kit for the Armed Forces DNA Identification Laboratory. Master of Science (Forensic Genetics), July 2004. 14 tables, 6 figures, references, 12 titles. To produce the most accurate and reliable results, forensic laboratories, such as AFDIL, must examine new technologies in the field of DNA analysis. The present study is the beginning of an internal validation of the most recent development in DNA quantitation, RT-PCR. The Quantifiler Human DNA Quantification Kit and Quantifiler Y Human Male DNA Identification Kit are RT-PCR assays that quantitate human and male DNA, and detect PCR inhibitors which may hinder the ability to produce a reliable STR profile. Sensitivity and non-probative case sample studies were performed according to the DAB guidelines. The Quantifiler kits were not as sensitive as had been previously reported by Applied Biosystems. The non-probative case sample study demonstrated results two fold greater than results from the Taqman® Alu-PCR Quantitation System, a RT-PCR assay developed and validated at AFDIL. At this time, it appears it may be in the best interest of AFDIL to continue its use of the Taqman® Alu-PCR Quantitation System.Item Validation and Optimization of Automated Instrumentation for Mitochondrial DNA Database Sample Processing(2004-06-01) Crispin, Shelley Jeanine; John Planz; Arthur Eisenberg; Joseph WarrenCrispin, Shelley Jeanine. Validation and Optimization of Automated Instrumentation for Mitochondrial DNA Database Sample Processing. Master of Science (Forensic Genetics), August 2004, 43 pages, 8 figures, 8 tables, 26 references. The validation of Qiagen 9604 BioRobot for DNA extraction and two Corbett CAS-1200 Liquid Precision Handling Systems for mitochondrial DNA database sample processing will alleviate sample backlogs and increase sample throughput. A manual cross-contamination study and an automated cross-contamination study showed that the use of automated instrumentation allowed for quicker and more accurate sample processing. The validation process was completed by processing a 96-well high throughput plate containing AFDIL employee samples. AFDIL loses approximately 10% of samples cycle sequenced using quarter volume BigDye terminator cycle reactions. To validate the use of half volume BigDye terminator cycled sequencing a 96-well high throughput plate containing AFDIL employee samples was used to compare the percentage of samples lost. On average 1.6% of the samples were lost per 96-well high throughput plate utilizing half volume BigDye terminator cycle sequencing reactions. This study has shown that the validation of half volume BigDye terminator cycle sequencing reactions will minimize the number of samples that will have to be reprocessed per 96-well high throughput plate.Item Validation of a Y-Chromosomal Short Tandem Repeat 10-Plex for Use in Fornesic and Paternity Laboratories(2002-08-01) Johnson, Cassie L.; Arthur Eisenberg; Joseph Warren; John PlanzJohnson, C., Validation of a Y-Chromosomal Short Tandem Repeat 10-Plex for Use in Forensic and Paternity Laboratories. Master of Science (Forensic Genetics), August, 2002. 94 pp., 6 tables, 7 figures, references, 21 titles. Y-chromosome microsatellite markers (Y-STRs) provide valuable information in cases of rape and questioned paternity, and they allow for the genetic identification of males. The present study validated a Y-STR 10-plex on the ABI Prism 3100 Genetic Analyzer for use in forensic and paternity laboratories at Orchid GeneScreen. Following optimization of the polymerase chain reaction, father-son pairs were analyzed to ensure that each pair generated identical haplotypes. The mutation rate varied between 0-0.238 (+/- 0.045, 95% confidence interval). The present study demonstrated that the 10-plex is sensitive to 0.75 ng and that female samples mixed with male samples did not interfere with Y-STR haplotyping. A population database of 525 males was developed and subsequently analyzed. Three instances of locus multiplication were observed, two at DYS19 and one at DYS435. Overall haplotype diversity was 0.996, suggesting that the 10-plex can efficiently distinguish male Y-STR profiles.Item Validation of Bodeplex 3 for Typing Small Amplicons and Low Copy Number DNA(2003-07-01) Chahrour, Maria H.; Arthur Eisenberg; John Planz; Joseph WarrenChahrour, Maria H., Validation of BodePlex3 for typing small amplicons and low copy number DNA. Master of Science (Forensice Genetics), July, 2003. 76 pp., 14 tables, 6 figures, references, 25 titles. The BodePlex 3 multiplex is a sensitive mini-STR typing system for analyzing highly degraded and low copy number (LCN) DNA. It allows for the typing of smaller amplicons and decreased quantities of DNA template, thus enhancing the sensitivity of STR analysis of compromised samples. The present study validated BodePlex 3 multiplex system on the ABI PRISM 3100 Genetic Analyzer to be used at The Bode Technology Group for forensic DNA analysis of small amplicons and LCN DNA. Validation experiments were performed according to the DNA Advisory Board (DAB) guidelines. Performance of the multiplex was accurate, reliable and reproducible. The results indicated that the typing system is highly specific for human DNA, sensitive for detecting profiles of LCN DNA, and is capable of resolving mixtures to a certain extent. In addition, this project outlined possible limitations that must be considered for successful use and interpretation of BodePlex 3 DNA profiling results.Item Validation of the Laboratory Information Systems Applications (LISA): Kinship Analysis Module(2008-11-01) Escobedo, IsraelEscobedo, Israel, Validation of the Laboratory Information Systems Applications (LISA): Kinship Analysis Module. Master of Science (Biomedical Sciences), December 2008, 155 pp., 10 tables, 21 figures, references, 34 titles. The Laboratory Information Systems Applications (LISA) is a LIMS system designed for the management and analysis of genetic data from large scale investigations of human identification. The validation study evaluated the pedigree construction and analytical tools of the Kinship Analysis module. The genetic data from old paternity cases was used to construct pedigrees under several scenarios that stimulate situations involving a missing child/parent. Each pedigree was analyzed to obtain a KI that measures the strength of the observed genetic evidence for an association made between a missing/deceased individual and a family reference pedigree to make identification. A common distribution of the number of observations per range of the 1n KI was observed in all scenarios. A concordance and reproducibility study was conducted for eight families using a validated kinship analytical program. LISA has the potential of becoming an efficient tool for human identification despite the limitations of the software.