Laborious NGS Library Prep: How to Ease Your NGS-Based HLA Typing?

The research field of HLA genotyping becomes revolutionized by Next Generation Sequencing (NGS). However, producing high-quality NGS libraries is labor-intensive and error prone. Read how to take advantage of NGS and to master the challenges of library prep…

The NGS revolution

High resolution typing of HLA (human leukocyte antigen) genes by NGS is becoming mainstream in high-throughput histocompatibility labs replacing the traditional Sanger sequencing. The advantages are obvious: NGS allows multiplexing of samples and provides a greater level of information in the highly polymorphic genomic HLA region. Determining the full sequence for class I and extended exon coverage for class II genes produces HLA types with far fewer ambiguities than traditional low-resolution methods like Sanger sequencing (1). In addition, NGS brings benefits of higher throughput and lower cost.

NGS library prep for HLA sequencing: The other side of the coin

Producing robust and high-quality DNA libraries from precious samples is a key requirement for HLA typing. However, a laborious and error-prone procedure. Users quickly suffer from an incredibly high number of pipetting steps and exhaustive long protocols. When was the last time you were annoyed by doing magnet bead-based cleanup steps in 96-well format or normalizations? As it is difficult to work concentrated for such a long time, the consequence is a high susceptibility to errors. This is topped with inter-user and inter-run variabilities and errors associated with manual pipetting affecting accuracy and reproducibility. Despite the loss of time, every failed sample costs a lot of money for reagents and consumables for NGS-library prep and sequencing itself. The loss of the sample material used can even be invaluable.

How to make tedious tasks a thing of the past

Wouldn't it be great to lower your workload but achieving a higher reproducibility? Automation is the solution!
A liquid handling robot translates a pipetting-intensive protocol into a ready-to-run method requiring minimal interventions and setup time. The automated workflow significantly reduces hands-on time, allows maximum walk-away time and relieves the extreme manual handling stress. Common error sources like inter-technician or inter-run variability and errors associated with manual pipetting are virtually eliminated. The results: More consistency and better accuracy. Significant time savings and the streamlined workflow help to increase your productivity. 

Benefits of automation at a glance:

  •  Significant reduction of hands-on time
  •  Elimination of common error sources 
  •  Increased reproducibility
  •  Less sample loss and reagent usage
  •  Relieve from manual handling stress

Proven reliability for HLA genotyping

Automated NGS sample prep methods have been qualified by kit manufacturers and customers to deliver sequencing results that are comparable or better to those from manual preparation. Accordingly, automated NGS library prep for the research of HLA typing (1,2,3) has been shown to save 30 percent time (2) compared to manual sample preparation whilst keeping the quality criteria yield, fragment length and sequencing concordance (2,3).

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