Evaluation of the AluQuant Human DNA Quantitation System Using the 96-Well Plate Format

Alexander, 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.