Strathprints Home | Open Access | Browse | Search | User area | Copyright | Help | Library Home | SUPrimo

Quantification of trace amounts of human and non-human mitochondrial DNA (mtDNA) using SYBR Green and real time PCR

Tobe, Shanan S. and Linacre, Adrian (2008) Quantification of trace amounts of human and non-human mitochondrial DNA (mtDNA) using SYBR Green and real time PCR. Forensic Science International: Genetics Supplement Series, 1 (1). pp. 71-73. ISSN 1875 1768

Full text not available in this repository. (Request a copy from the Strathclyde author)

Abstract

There are currently no tests available to quantify total non-human mammalian mtDNA. Standard universal DNA quantification tests are unsuitable due to the large size difference between nuclear and mitochondrial genomes and the ubiquity of human mtDNA. A method has therefore been developed to quantify total mammalian mtDNA and total human mtDNA present in a sample using SYBR Green. Mammalian primers designed to react with all mammals were designed on the 12S and human specific primers were designed on the cytochrome b gene. Each primer set was reacted separately with sample and SYBR Green and detected using RT-PCR. A standard curve was developed using dilutions ranging from 1 billion copies to 100 copies of mtDNA. Twenty-four human samples were analysed and an average log (copy number) human/universal ratio of 1.00 was obtained. Samples falling below this ratio will contain some non-human mtDNA while samples falling above this ratio contain human mtDNA only. Twenty-nine mammal samples were also tested. 96.6% of these showed human contamination to some extent. This test is able to quantify mtDNA down to the femtogramme (10E−15g) level.

Item type: Article
ID code: 33829
Keywords: quantification, human mtDNA, non-human mtDNA, human contamination, Genetics, Genetics, Pathology and Forensic Medicine
Subjects: Science > Natural history > Genetics
Department: Faculty of Science > Pure and Applied Chemistry
Related URLs:
    Depositing user: Pure Administrator
    Date Deposited: 28 Sep 2011 14:00
    Last modified: 05 Sep 2014 11:17
    URI: http://strathprints.strath.ac.uk/id/eprint/33829

    Actions (login required)

    View Item