GenDx NGSgo HLA typing

High resolution HLA typing data is critical for histocompatibility testing. NGS technology provides allele-level resolution and supports high throughput workflows.

GenDx has pioneered the development of software and reagents – such as NGSgo® – for high resolution HLA typing using NGS.

Automation of NGSgo HLA typing

Automating the NGSgo workflow on the Freedom EVO NGS workstation minimizes the risk of cross-contamination and increases the reproducibility of library preparation for up to 96 samples.

The platform uses advanced air displacement technology to enable precise eight-channel pipetting.

The instrument also includes:

  • three Inheco CPAC thermal devices to keep reagents cool and ensure optimal temperatures for each incubation step
  • an Inheco Thermoshake heated shaker
  • a 96-position magnetic plate separator
  • a Robotic Manipulator Arm™ for efficient bed clean-up
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Simplified Workflow

The following protocol steps are automated on the Freedom EVO NGS workstation: generation of amplicon pooling, fragmentation and adapter ligation, bead clean-ups and size selection, indexing PCR and library pooling. The quantification, normalization and indexing can be performed in an offline thermocycler, or fully automated by integrating a Tecan microplate reader and thermocycler onto the work deck, significantly increasing walkaway time.


Automation effectiveness in HLA typing

Automating the NGSgo library preparation workflow on the Freedom EVO NGS workstation generates high yield libraries. After sequencing, the number of reads assigned to each library was within 17 percent of the average. GenDx NGSengine® software generated HLA typing results for all samples and loci, demonstrating the effectiveness of this automated workflow for library preparation.

Sequencing metrics confirmed the generation of high quality libraries:

  • sample mappability was over 85 percent for all samples
  • locus mappability exceeded 90 percent
  • all loci were fully covered with a read depth of at least 194 in the HLA core sequencing regions
  • heterozygous positions were well balanced and cleanly separated from the sequencing noise


For research use only.