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Development of Publicly Available Forensic DNA Sequence Mixture Data.

Erica L Romsos¹, Kevin M Kiesler¹, Carolyn R Steffen¹

¹National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, MD 20899, USA.

|March 28, 2025

Summary

This summary is machine-generated.

Forensic scientists developed complex DNA mixture samples for next-generation sequencing (NGS) analysis. These samples aid in validating probabilistic genotyping software for improved forensic casework interpretation.

Keywords:

bioinformatics forensic DNA mixtures sequencing training data validation

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Area of Science:

Forensic Science
Genetics
Bioinformatics

Background:

The Scientific Working Group on DNA Analysis (SWGDAM) identified data needs for sequence-based probabilistic genotyping software in 2018.
Engagement with developers highlighted requirements for comprehensive mixture sample datasets.

Purpose of the Study:

To create a diverse set of DNA mixture samples representing forensic casework variability.
To support the development and validation of next-generation sequencing (NGS) based probabilistic genotyping tools.

Main Methods:

74 mixture samples were generated from 11 single-source samples.
Samples included varying numbers of contributors (3-5), DNA input levels, and degradation.
Allelic overlap was evaluated to select representative sample combinations.

Main Results:

A 96-well plate design was utilized for sample distribution.
Included are triplicates of 3-person mixtures (1-5% minor components), degraded DNA mixtures, and 4-5 person mixtures.
A single-source dilution series was also included for sensitivity testing.

Conclusions:

The prepared mixture samples have been sequenced using commercial forensic STR and SNP kits.
FASTQ data files and metadata are publicly available for further research and software development.