NGS Workflow

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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 0.8µg of DNA required for 11 locus amplification
  • Compatible with multiple DNA sources: whole blood, cord blood, buccal swabs & saliva
  • 5, 7 or 11 locus amplifications
  • Scalable up to 96 samples / 11 loci, up to 192 samples / 7 loci and 288 / 5 loci***

Library Preparation

  • 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

 

Sequencing - Illumina Sequencers

  • MiniSeq, MiSeq, iSeq 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
  • Less than 5 minutes analysis time per 11 loci sample with recommended specifications
  • Two independent genotyping algorithms
  • 24 Quality Control Metrics for confident assignment
  • 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


All timings are for 24 sample, Holotype HLA v3.0
*     Timings are based on the liquid handler used
**   On a MiSeq 300-cycle Micro flow cell
*** On a 300-cycle Standard flow cell with cluster density ~1,000K/mm²

The workflow is available as Research Use Only. Details about the CE-IVD workflow is available upon request.