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Y-STR mixture deconvolution by single-cell analysis.

Kaitlin Huffman1, Erin Hanson1,2,3, Jack Ballantyne1,2,3

  • 1Graduate Program in Chemistry, Department of Chemistry, University of Central Florida, Orlando, Florida, USA.

Journal of Forensic Sciences
|October 2, 2022
PubMed
Summary
This summary is machine-generated.

Direct single-cell genotyping effectively deconvolutes challenging Y-STR DNA mixtures from multiple male contributors. This technique isolates individual cells for analysis, simplifying interpretation in forensic casework involving complex DNA samples.

Keywords:
DSCSY-STR analysismixture deconvolutionphysical micromanipulationsingle-cell analysistouch DNA

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Area of Science:

  • Forensic Science
  • Genetics
  • Molecular Biology

Background:

  • Y-STR typing simplifies DNA mixtures with female DNA but struggles with multiple male contributors.
  • Current probabilistic genotyping software is not validated for complex Y-STR mixture analysis.
  • Single-cell genotyping offers a potential solution for deconvoluting mixed Y-STR profiles.

Purpose of the Study:

  • To develop and validate a direct single-cell genotyping method for Y-STR mixture analysis.
  • To assess the efficacy of physical micromanipulation and direct PCR for isolating and analyzing single cells from complex mixtures.
  • To apply the technique to buccal cell mixtures and a mock forensic case involving touch DNA.

Main Methods:

  • Physical micromanipulation using a tungsten needle to isolate single cells.
  • Direct PCR with optimized reaction volume and cycle number.
  • Application to equimolar 2- and 3-person male buccal cell DNA mixtures.
  • Analysis of a mock touch DNA scenario involving consecutive handgun firing by two males.

Main Results:

  • Successfully deconvoluted 2- and 3-person male buccal cell mixtures into individual donor Y-STR haplotypes.
  • Recovered single-source Y-STR profiles from both individuals in the handgun trigger touch DNA scenario.
  • Identified specific DNA profiles from the gun's trigger and handle, differentiating between individuals.

Conclusions:

  • Direct single-cell subsampling is a promising and simple method for analyzing and interpreting Y-STR mixtures.
  • This technique can overcome limitations of current Y-STR mixture analysis methods.
  • Potential applications in forensic science for complex DNA mixture interpretation.