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Automating the Illumina DNA library preparation kit for whole genome sequencing applications on the flowbot ONE liquid handler robot

  • 0Department of Medical Microbiology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands.
Scientific Reports +

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April 8, 2024

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April 8, 2024

View abstract on PubMed

Summary

This summary is machine-generated.

Automating whole-genome sequencing library preparations (LP) on the flowbot ONE robot significantly reduces hands-on time (HOT) and improves reproducibility. This automated workflow ensures comparable results to manual methods, enhancing clinical diagnostic efficiency.

Area Of Science

  • Genomics
  • Molecular Biology
  • Clinical Diagnostics

Background

  • Whole-genome sequencing (WGS) is transitioning to routine clinical diagnostics.
  • Library preparation (LP) is a critical, labor-intensive step in WGS.
  • Manual LP is prone to human error and requires substantial hands-on time (HOT).

Purpose Of The Study

  • To automate the Illumina DNA Prep library preparation workflow using the flowbot ONE robot.
  • To minimize human error and reduce hands-on time (HOT) in WGS library preparation.
  • To assess the reliability, reproducibility, and efficiency of the automated LP workflow.

Main Methods

  • The flowbot ONE robot was programmed and optimized for automated Illumina DNA Prep.
  • 16 library preparations were performed manually and automatically for comparison.
  • Reproducibility was assessed by re-processing eight library preparations using the automated workflow.

Main Results

  • Automated and manual LPs showed comparable library DNA yields (median 1.5-fold difference).
  • Assembly quality values were similar between automated and manual methods.
  • 100% concordance was observed in core genome multilocus sequence typing results.
  • Hands-on time (HOT) was reduced from 125 minutes (manual, 8 LPs) to 25 minutes (automated, 8 LPs).
  • Turn-around time was reduced for the automated workflow (170 min) compared to manual (200 min).

Conclusions

  • The automated flowbot ONE workflow provides consistent and reproducible results for WGS library preparation.
  • Automation significantly reduces hands-on time (HOT) and turn-around time compared to manual methods.
  • Automated LP is a reliable alternative for clinical diagnostic WGS workflows.