This webinar provides a practical approach to using reference standards to define the performance characteristics of new next-generation sequencing assays during validation.

In this webinar, Robyn Sussman and Ashkan Bigdeli from the Center for Personalized Diagnostics at the University of Pennsylvania discuss how they used reference standards as part of the validation workflow for their NGS assays.

In one case, they used standards with varying degrees of damage to validate a low-input assay that can accommodate highly compromised formalin-fixed, paraffin embedded tissues.

Sussman and Bigdeli validated a small, amplicon-based sequencing panel to accommodate the low-yield and low-quality specimens that do not qualify for larger sequencing panels. The team used Horizon’s Quantitative Multiplex Reference Standard gDNA along with Quantitative Multiplex Formalin Compromised DNA (Mild, Moderate and Severe) to assess the input requirements for the assay and the performance characteristics of samples with varying DNA quality.

For larger panels with whole coding sequencing coverage, copy number changes to specific genes can be predicted algorithmically. The University of Pennsylvania team will also discuss how they validated a copy number caller for a solid tumor sequencing panel. For this project, they used Horizon’s Structural Mutliplex Reference Standard gDNA and Structural Multiplex FFPE Reference Standard to define the performance characteristics of the caller, which will be used for both fresh and paraffin-embedded tissue.