NGS Workflow

Check out our NGS workflow!

Holotype HLA is an NGS-based HLA typing workflow that is fully automated for multiple liquid handling robots from sample prep, library prep and sequencing on any Illumina® sequencer with automatic single-pass data analysis with HLA Twin.

Sample Preparation

  • Less than 1μg of DNA required for 11 locus amplification
  • Successfully amplified whole blood, cord blood, buccal swabs & saliva

Library Preparation

  • 5, 7 or 11 locus amplifications
  • Scalable up to 72 samples / 11 loci, up to 128 samples / 7 loci and 188 / 5 loci***
  • 4 hours hands-on time for 24 samples
  • Less than 48 hours turnaround time from sample to result
  • Library Size Selection: Pippin Prep or Magnetic Beads
  • Holotype libraries can be combined with other libraries for optimized sequencing cost

All timings are for 24 sample, Holotype HLA V2.2
*     Timings are based on the Biomek 4000 by Bechman Coulter.

**   On a MiSeq 300-cycle Micro flow cell
*** On a 300-cycle Standard flow cell with cluster density ~1,000K/mm²

Sequencing - Illumina Sequencers

  • MiniSeq, MiSeq and NextSeq supported
  • Any size flow cell supported depending on throughput
  • 300- and 500-cycle chemistry supported

Data Analysis with HLA Twin

  • Fully automated analysis after Illumina sequencing run
  • Two independent genotyping algorithms in HLA Twin for orthogonal validation
  • 23 Quality Control Metrics for confident assignment
  • Support for Mac, Windows and Linux, standalone and server-client setups
  • Balanced coverage between alleles at a locus, between loci in a sample and among samples
  • Deep and even coverage across the whole length of the targeted region
  • Integration with LIMS, such as Histotrac
The workflow is available as Research Use Only. Details about the CE-IVD workflow is available upon request.