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Updated: Oct 18, 2025

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Pushing the Boundaries: Forensic DNA Phenotyping Challenged by Single-Cell Sequencing.

Marta Diepenbroek1, Birgit Bayer1, Katja Anslinger1

  • 1Department of Forensic Genetics, Institute of Legal Medicine, Ludwig Maximilian University of Munich, Nußbaumstraße 26, 80336 Munich, Germany.

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|September 28, 2021
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Summary

Forensic DNA phenotyping can now analyze mixed samples using single-cell sequencing. This advanced technique successfully deconvolutes complex DNA mixtures, improving forensic investigations.

Keywords:
DEPArrayFDPHIrisPlex-Sancestry predictionforensic DNA phenotypinglow template DNAltDNAmassively parallel sequencingmixture deconvolutionnext-generation sequencingphenotype predictionsingle-cell genomicssingle-cell sequencing

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Area of Science:

  • Forensic Science
  • Genomics
  • Molecular Biology

Background:

  • Single-cell sequencing is emerging in forensics for mixture deconvolution, particularly for same-cell-type traces.
  • Current forensic DNA phenotyping methods are limited with mixed samples.

Purpose of the Study:

  • To evaluate the efficacy of single-cell sequencing for forensic DNA phenotyping.
  • To assess the potential of this method in deconvoluting mixed DNA traces for phenotype and ancestry analysis.

Main Methods:

  • White blood cells were isolated from blood samples using the DEPArray™ N×T System.
  • DNA libraries were prepared using the Ion AmpliSeq™ PhenoTrivium Panel for SNP analysis.
  • Groups of single cells (20, 10, 5) and individual cells were sequenced.

Main Results:

  • Nearly complete consensus profiles were achieved for both cell groups and single cells.
  • Accurate phenotype (eye, hair, skin color) and ancestry predictions were obtained.
  • Re-amplification of single-cell libraries also yielded correct predictions.

Conclusions:

  • Single-cell sequencing is a promising tool for deconvoluting mixed forensic DNA traces.
  • This technology enhances the capabilities of forensic DNA phenotyping.
  • It offers potential for improved analysis of complex forensic samples.