uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
Rapid quantification and sex determination of forensic evidence materials
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Genomics.
2003 (English)In: Journal of Forensic Sciences, ISSN 0022-1198, E-ISSN 1556-4029, Vol. 48, no 6, 1280-1287 p.Article in journal (Refereed) Published
Abstract [en]

DNA quantification of forensic evidence is very valuable for an optimal use of the available biological material. Moreover, sex determination is of great importance as additional information in criminal investigations as well as in identification of missing persons, no suspect cases, and ancient DNA studies. While routine forensic DNA analysis based on short tandem repeat markers includes a marker for sex determination, analysis of samples containing scarce amounts of DNA is often based on mitochondrial DNA, and sex determination is not performed. In order to allow quantification and simultaneous sex determination on minute amounts of DNA, an assay based on real-time PCR analysis of a marker within the human amelogenin gene has been developed. The sex determination is based on melting curve analysis, while an externally standardized kinetic analysis allows quantification of the nuclear DNA copy number in the sample. This real-time DNA quantification assay has proven to be highly sensitive, enabling quantification of single DNA copies. Although certain limitations were apparent, the system is a rapid, cost-effective, and flexible assay for analysis of forensic casework samples.

Place, publisher, year, edition, pages
2003. Vol. 48, no 6, 1280-1287 p.
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-92986PubMedID: 14640271OAI: oai:DiVA.org:uu-92986DiVA: diva2:166326
Available from: 2005-04-29 Created: 2005-04-29 Last updated: 2011-10-26Bibliographically approved
In thesis
1. Sensitive Forensic DNA Analysis: Application of Pyrosequencing and Real-time PCR Quantification
Open this publication in new window or tab >>Sensitive Forensic DNA Analysis: Application of Pyrosequencing and Real-time PCR Quantification
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The field of forensic genetics is growing fast and the development and optimisation of more sensitive, faster and more discriminating forensic DNA analysis methods is highly important. In this thesis, an evaluation of the use of novel DNA technologies and the development of specific applications for use in forensic casework investigations are presented.

In order to maximise the use of valuable limited DNA samples, a fast and user-friendly Real-time PCR quantification assay, of nuclear and mitochondrial DNA copies, was developed. The system is based on the 5’ exonuclease detection assay and was evaluated and successfully used for quantification of a number of different evidence material types commonly found on crime scenes. Furthermore, a system is described that allows both nuclear DNA quantification and sex determination in limited samples, based on intercalation of the SYBR Green dye to double stranded DNA.

To enable highly sensitive DNA analysis, Pyrosequencing of short stretches of mitochondrial DNA was developed. The system covers both control region and coding region variation, thus providing increased discrimination power for mitochondrial DNA analysis. Finally, due to the lack of optimal assays for quantification of mitochondrial DNA mixture, an alternative use of the Pyrosequencing system was developed. This assay allows precise ratio quantification of mitochondrial DNA in samples showing contribution from more than one individual.

In conclusion, the development of optimised forensic DNA analysis methods in this thesis provides several novel quantification assays and increased knowledge of typical DNA amounts in various forensic samples. The new, fast and sensitive mitochondrial DNA Pyrosequencing assay was developed and has the potential for increased discrimination power.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2005. 44 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 33
Genetics, forensic, sensitive DNA analysis, DNA quantification, mtDNA, Real-time PCR, Pyrosequencing, Genetik
National Category
Medical Genetics
urn:nbn:se:uu:diva-5775 (URN)91-554-6234-0 (ISBN)
Public defence
2005-05-21, Rudbecksalen, Rudbecklaboratoriet, Uppsala, 09:00
Available from: 2005-04-29 Created: 2005-04-29Bibliographically approved

Open Access in DiVA

No full text


Search in DiVA

By author/editor
Allen, Marie
By organisation
Department of Immunology, Genetics and PathologyGenomics
In the same journal
Journal of Forensic Sciences
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 306 hits
ReferencesLink to record
Permanent link

Direct link