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Quantifiler Observations of Relevance to Forensic Casework

NCJ Number
222159
Journal
Journal of Forensic Sciences Volume: 53 Issue: 1 Dated: January 2008 Pages: 135-141
Author(s)
Irene Koukoulas Ph.D.; Fiona E. O'Toole B.Sc.; Peta Stringer Ph.D.; Roland A.H. van Oorschot Ph.D.
Date Published
January 2008
Length
7 pages
Annotation
This report on in-house validation studies of the Quantifiler (QF) kit (Victoria Police Forensic Services Centre [Australia]) focused on DNA concentration estimation differences between alternative quantification methods, concentration differences between DNA standards from the same supplier as well as between different suppliers, the effectiveness of the internal PCR control (IPC) system, the effect of high starting DNA template concentration on the IPC, and quantification results when using alternative analysis settings with the 7500 SDS software (v1.3).
Abstract
The results show that if the Promega K562 DNA standard (system lot number 206778) and any further lot numbers of the same DNA concentration are used in conjunction with the QF kit, acceptable peak heights can be achieved in subsequent Profiler Plus amplifications of 1 ng of sample DNA for both homozygote and heterozygote alleles in Profiler Plus amplification and potentially other STR systems. This is not the case when QF standard lot A and any other lots with the same DNA concentration are used. The validation studies have also shown a two-fold difference in DNA concentration between lots A and B of the standard provided in the QF kit. Of the four purification methods studied, the QIAquick method was the most successful. The use of QF to quantify DNA for down-stream processing of forensic casework samples has improved the ability to more accurately determine and predict STR amplification success. It can also estimate a wider range of DNA concentration, and it uses less DNA while reducing user time and effort compared to other quantification methods. Detailed descriptions are provided for materials and methods used in the validation studies. 4 tables, 4 figures, and 19 references