Browsing by Subject "PCR inhibitors"
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Item A Bone and Buccal Sensitivity Study Comparison and Stability Study using the PowerPlex[R] Fusion 6C System(2018-05) McDaniel, Ethan L.; Warren, Joseph E.; Planz, John V.; Krishnamoorthy, Raghu R.; Gaydosh-Combs, LauraA validation study, a bone sensitivity study and a stability study were performed using the PowerPlex[R] Fusion 6C System. These studies were performed on a 7500 Real-Time PCR System, 9700 GeneAmp Thermocycler and 3500xL Genetic Analyzer. Buccal DNA was used to develop a method to analyze the DNA profiles gathered during the bone sensitivity study and stability study. DNA profiles for specific concentrations of DNA in solution were obtained during the bone sensitivity study. The stability study showed profiles exhibiting the effects of metal PCR inhibitors being introduced to the DNA extract solutions. Full profiles were obtained for calcium concentrations less than 7.35 mM, while the instrument was fully inhibited for copper concentrations between 0mM and 7.35 mM. Based on the limited data, the PowerPlex Fusion 6C System cannot tolerate copper when present in DNA solutions; whereas, calcium may be tolerated as an inhibitor up to 7.35mM.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.