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Validation of Reduced‐Scale Reactions for the Quantifiler™ Human DNA Kit
Author(s) -
Westring Christian G.,
Kristinsson Richard,
Gilbert Dustin M.,
Danielson Phillip B.
Publication year - 2007
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2007.00525.x
Subject(s) - scale (ratio) , dna , computational biology , computer science , forensic engineering , engineering , biology , genetics , geography , cartography
Accurate quantification of DNA samples is an important step in obtaining accurate and reproducible short tandem repeat (STR) profiles. Quantitative real‐time‐PCR has improved the speed and accuracy of DNA quantification over earlier methods, albeit at significantly greater cost per reaction. Here, the performance of reduced volume (10 μL) DNA quantification assays using the Quantifiler™ Human DNA Quantification Kit was evaluated using commercial standards and single source biological stains (e.g., venous blood, saliva, and semen). In addition, casework‐type samples including those subjected to environmental contaminants containing PCR inhibitors and samples having undergone extensive DNA degradation were also quantified. The concentration of DNA in various forensic samples ranged from 0 to 2.9 ng/μL depending on sample source and/or environmental insult. Compared to full‐scale reactions, reduced volume assays displayed equivalent to improved amplification efficiency and sample‐to‐sample reproducibility (±0.01–0.17 C T FAM ). Furthermore, the use of data from reduced‐scale Quantifiler™ reactions facilitated the accurate determination of the amount of sample DNA extract needed to generate quality STR profiles. The use of 10 μL‐scale Quantifiler™ reaction volumes has the practical benefit of increasing the effective number of reactions per kit by 250%; thereby reducing the cost per assay by 60% while consuming less sample. This is particularly advantageous in cases of consumptive testing.