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Assessing non-LUS stutter in DNA sequence data.

Olivia D'Angelo¹, Amber C W Vandepoele¹, Jonathan Adelman¹

¹Forensic & National Security Sciences Institute, Syracuse University, 100 College Place 120 Life Science Building, Syracuse, NY 13244, USA.

Forensic Science International. Genetics

|April 23, 2022

Summary

This summary is machine-generated.

Forensic DNA analysis using massively parallel sequencing (MPS) can now better characterize stutter artifacts. This study reveals that analyzing motif- and allele-specific stutter, including non-longest uninterrupted stretch (LUS) stutter, improves accuracy in forensic DNA profiling.

Keywords:

DNA sequence LUS Massively parallel sequencing (MPS)Non-LUS Short tandem repeats (STR)Stutter

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Area of Science:

Forensic Science
Molecular Biology
Genetics

Background:

Forensic DNA analysis relies on short tandem repeats (STRs) for individual identification.
Stutter, a PCR-induced artifact, challenges accurate DNA profile interpretation, especially in mixtures.
Traditional capillary electrophoresis (CE) limits understanding of stutter due to size-based separation.

Purpose of the Study:

To characterize allele- and motif-specific stutter using massively parallel sequencing (MPS) data.
To investigate the impact of non-longest uninterrupted stretch (non-LUS) stutter on forensic DNA analysis.
To develop more accurate methods for differentiating stutter artifacts from true alleles.

Main Methods:

Analysis of stutter sequences from 539 samples amplified with ForenSeq and PowerSeq 46GY kits.
Sequencing performed on the Illumina MiSeq FGx platform.
Quantification and characterization of both LUS and non-LUS stutter products at locus and allele levels.

Main Results:

Massively parallel sequencing (MPS) data allows for detailed characterization of stutter, including non-LUS stutter.
While LUS stutter occurs at higher rates, non-LUS stutter products are detectable and can influence mixture analysis.
Allele- and motif-specific stutter models provide more accurate differentiation of stutter from true alleles.

Conclusions:

Characterizing motif- and allele-specific stutter, including non-LUS variants, enhances the accuracy of forensic DNA profiling.
MPS-based stutter analysis offers a more comprehensive understanding compared to traditional CE methods.
This approach is crucial for accurate analysis of complex DNA mixtures, particularly those with low-level contributors.