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RMplex: An efficient method for analyzing 30 Y-STRs with high mutation rates.

Arwin Ralf1, Dion Zandstra1, Natalie Weiler2

  • 1Department of Genetic Identification, Erasmus MC, University Medical Center Rotterdam, Rotterdam, the Netherlands.

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|September 20, 2021
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Summary

Rapidly mutating Y-STRs (RM Y-STRs) offer higher resolution for differentiating paternal lineages. The new RMplex system analyzes 30 RM Y-STRs, aiding criminal casework with challenging DNA mixtures and familial searching.

Keywords:
Forensic validationGenetic identificationMPSMale differentiationMassively parallel sequencingMultiplex PCRRM Y-STRsRapidly mutating Y-STRsRelative separationY-STRs

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Area of Science:

  • Forensic Genetics
  • Human Genetics
  • Molecular Biology

Background:

  • Standard Y-STRs have limitations in differentiating closely related males.
  • Rapidly mutating Y-STRs (RM Y-STRs) provide higher resolution for paternal lineage analysis.
  • Recent discoveries have expanded the number of known RM Y-STRs.

Purpose of the Study:

  • To develop and validate RMplex, a genotyping system for 30 high-mutation-rate Y-STRs.
  • To enable higher-resolution differentiation of paternally related men.
  • To enhance forensic applications in criminal casework.

Main Methods:

  • Developed RMplex for analyzing 30 RM Y-STRs (44 male-specific loci total) using multiplex PCR.
  • Utilized capillary electrophoresis (CE) and massively parallel sequencing (MPS) for genotyping.
  • Validated the system for repeatability, reproducibility, low DNA input, and performance with inhibitors and female DNA.

Main Results:

  • RMplex efficiently genotypes 30 RM Y-STRs, including all currently known ones.
  • Validation demonstrated repeatable and reproducible results with low DNA input (100-250 pg).
  • The system performed well with inhibitors and excess female DNA, and successfully differentiated males within pedigrees.

Conclusions:

  • RMplex is an efficient genotyping system for high-resolution paternal lineage analysis.
  • The system shows promise for criminal casework, especially in sexual assault cases and familial searching.
  • Further forensic validation is expected to establish RMplex in routine forensic applications.