Internal Validation of Quantifiler Human Quantfilier Y Human male DNA Quantification Kits Using Applied Biosystems 7500 Real-Time PCR

Jones, Brekeitrea Minyon. Internal Validation of Quantifiler Human and Quantifiler Y Human DNA Quantification kits using the Applied Biosystems 7500 Real-Time PCR System. Master of Science (Forensic Genetics), August 2004, 36 pages, 4 figures, 8 tables, 18 references. Quantitation of DNA is an essential step in DNA testing and methods used to quantitate DNA must be human specific. Quantitation of DNA is also important because the PCR reaction performs optimally in a narrow concentration range. Applied Biosystems (Foster City, CA) has recently introduced two new DNA quantitation kits, Quantifiler Human and Quantifiler Y Human Male DNA Quantification kits, which quantify the total amount of amplifiable DNA present in a sample. Both kits are based on the 5’ nuclease assay and Taqman probe-based technology which allow for a more accurate result which will minimize further problems downstream. To validate the use of the Quantifiler kits, a series of studies were performed to determine the reliability and performance of the kits. Results demonstrate this quantitation method produces similar results to those obtained by the Quantiblot method. In addition, results demonstrate that this quantitation method is highly sensitive and yields consistent results.