Abstract
A modified alkaline lysis protocol for extracting DNA from forensically relevant specimens is evaluated and compared with the chelex 100 method. For whole blood, bloodstains and sperm stains, both methods yielded comparable results after amplification for a pentameric STR locus (HumCD4). The main advantages of the new method are: only approximately ten minutes and two pipetting steps are necessary and the expenses for the extraction are extremely low as only NaOH, TrisHC1 buffer and a single microcentrifuge tube are required. Alkaline lysis also proved to yield DNA suitable for typing longer STRs by using dye-labeled primers and capillary electrophoresis. These advantages should render this protocol especially interesting for high-throughput laboratories in combination with multiplex PCR and fluorescent dye technology, although the storability of the extracts proved to be problematic.
Keywords:
alkaline lysis, chelex 100, DNA typing, extraction, forensic science, HumCD4, short tandem repeats
Paper ID: JFS45326
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Author Paffumi, E Affiliation Institute for Energy, European Commission, DG-JRC, Petten, ZG, Netherlands and Materials Research Centre, School of Engineering, University of Wales, Petten, ZG, UK
Author Nilsson, K Affiliation Institute for Energy, European Commission, DG-JRC, Petten, ZG, Netherlands
Author Taylor, N Affiliation Institute for Energy, European Commission, DG-JRC, Petten, ZG, Netherlands
Author Hurst, R Affiliation Institute for Energy, European Commission, DG-JRC, Petten, ZG, Netherlands
Author Bache, M Affiliation Materials Research Centre, School of Engineering, University of Wales, Petten, ZG, UK
Author Klintschar M, Neuhuber F
Title Evaluation of an alkaline lysis method for the extraction of DNA from whole blood and forensic stains for STR analysis
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