The ForenSeq MainstAY Workflow

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Next-generation sequencing (NGS) platforms emerged as an alternative to capillary electrophoresis (CE) nearly two decades ago. The ability to generate significantly more information from a DNA sample, coupled with decreasing costs, has transformed the field of genetics, and more recently, human identification. Global forensic laboratories have started evaluating how NGS can provide additional capabilities to help advance investigations, such as whole mitochondrial sequencing, externally visible phenotype estimation, mRNA expression, and paternity and kinship analyses. In addition to providing new tools, NGS provides a more powerful look at the foundational marker of human ID, short tandem repeats (STRs). Unlike traditional length-based analyses, sequencing STRs uncovers more information from the same DNA samples, identifying previously unknown alleles, characterizing sequence-level variation in core forensic loci, and providing information that can be leveraged in mixture deconvolution.

To provide forensic laboratories more ways to maximize sample information, Verogen introduces the ForenSeq MainstAY workflow— a targeted sequencing approach for the core autosomal and Y loci that are part of CODIS, the European Standard Set (ESS), and the YHRD (Y Chromosome Haplotype Reference Database), optimized for the MiSeq FGx Sequencing System. By focusing on the common STR markers needed for human identification, using NGS for routine casework and confirmatory testing is simple and easy.

In this presentation, Verogen Staff Scientist Dr. Bobby LaRue will:

• Discuss the benefits of NGS over CE analysis in terms of sequence context, the ability to multiplex more markers, and the analysis of degraded samples which may not generate full profiles with CE
• Share technical details of the ForenSeq MainstAY Kit and examine performance with challenging mock casework samples
• Present case studies that compare the merits and limitations of CE and NGS in forensic casework
• Highlight future capabilities for the Verogen STR portfolio