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Massively parallel sequencing of forensically relevant single nucleotide polymorphisms using TruSeq™ forensic

David H Warshauer1, Carey P Davis, Cydne Holt

  • 1Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX, 76107, USA.

International Journal of Legal Medicine
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The TruSeq™ Forensic Amplicon protocol effectively prepares DNA for massively parallel sequencing (MPS). This method accurately detects forensic genetic markers, including single nucleotide polymorphisms (SNPs), with high concordance across different labs and whole genome sequencing.

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Area of Science:

  • Forensic genetics
  • Molecular biology
  • Genomics

Background:

  • The TruSeq™ ChIP-Seq protocol was adapted for forensic applications.
  • Targeted sequencing requires efficient library preparation methods.

Purpose of the Study:

  • To evaluate the TruSeq™ Forensic Amplicon library preparation protocol for detecting forensic genetic markers.
  • To assess the concordance of genotype calls with other methods.

Main Methods:

  • The TruSeq™ Forensic Amplicon protocol was used to prepare libraries from 12 reference samples.
  • 160 single nucleotide polymorphisms (SNPs), including human identification SNPs (iSNPs), ancestry, and phenotypic SNPs (apSNPs), were targeted.
  • Results were compared to a second laboratory and whole genome sequencing (WGS).

Main Results:

  • High concordance was observed for genotype calls generated by the TruSeq™ Forensic Amplicon protocol.
  • The protocol demonstrated effectiveness in preparing amplified nuclear DNA for massively parallel sequencing (MPS).

Conclusions:

  • The TruSeq™ Forensic Amplicon protocol is a sensitive and effective method for forensic DNA analysis.
  • This protocol facilitates the targeted sequencing of forensic genetic markers for human identification, ancestry, and phenotypic analysis.