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DNA profiling of single sperm cells after whole genome amplification.


Glenn M.G. Theunissen

Andrew Gibb

Burkhard Rolf

Sophia Forat



Intimate swabs taken for examination in sexual assault cases typically yield mixtures of sperm and epithelial cell types. While powerful, differential extraction protocols to overcome such cell type mixtures by separate lysis of epithelial cells and spermatozoa can still prove ineffective, in particular if only few sperm cells are present or if swabs contain sperm from more than one individual leading to complex low level DNA mixtures. A means to avoid such mixtures consists in the analysis of single micromanipulated sperm cells. However, the quantity of DNA from single sperm cells is not sufficient for conventional STR analysis. Here, we describe a simple method for micromanipulating individual sperm cells from intimate swabs and show that whole genome amplification can generate sufficient amounts of DNA from single cells for subsequent DNA profiling. We recovered over 80% of alleles of haploid autosomal STR profiles from the majority of individual sperm cells. Furthermore, we demonstrate that in mixtures of sperm from two contributors, Y-STR and X-STR profiles of individual sperm cells can be used to sort the haploid autosomal profiles to develop the diploid consensus STR profiles of the individual donors. Finally, by analyzing single sperm cells from mock sexual assault swabs with one or two sperm donors, we showed that our protocols enabled the identification of the unknown male contributors.


THEUNISSEN, G.M.G., GIBB, A., KONG THOO LIN, P., ROLF, B., FORAT, S. and JÄGER, R. 2021. DNA profiling of single sperm cells after whole genome amplification. Forensic science international: reports [online], 4, article 100240. Available from:

Journal Article Type Article
Acceptance Date Oct 13, 2021
Online Publication Date Oct 16, 2021
Publication Date Nov 30, 2021
Deposit Date Oct 22, 2021
Publicly Available Date Oct 22, 2021
Journal Forensic Science International: Reports
Print ISSN 2665-9107
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 4
Article Number 100240
Keywords Short tandem repeat (STR); Forensic genetics; DNA typing; Whole genome amplification; Y-STR; X-STR; Micromanipulation; Sperm cell; Sexual assault
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