Watch this webinar on demand: Volta + IDT Precision at Scale - Revolutionizing Genomic Sample Preparation

On January 14, 2025, Volta Labs and IDT partnered to produce this webinar presented by Abdul Mohammed, Head of Applications Development at Volta Labs, and Nick Downey, NGS Collaborations Lead at IDT. They discuss performance data from Volta's IDT xGen™ NGS Hybridization Capture app on the Callisto Sample Prep System and how Callisto addresses existing challenges to offer an innovative and reliable solution for automating hybridization capture workflows.

To evaluate Volta's IDT xGen™ NGS Hyb Capture app performance on Callisto, three experiments were performed in this study. Phase 1 and 2 evaluate inter-user variability of single-plex libraries of the Exome v2 Hyb panel and the Pan-Cancer Hyb panel, respectively, compared directly to manually prepared libraries. Phase 3 evaluates multiplexing on Callisto, from 1-plex all the way up to 20-plex libraries. 

Phase 1: Operator variability with large panels using the xGen IDT Exome V2 panel, comparing directly against manual preparations

• 100 ng HG002 prepared using the xGen™ DNA Lib Prep EZ for exome, xGen Exome Hyb Panel v2
• This panel has approximately 415K individual probes spanning 34 Mb of target region for the human exome
• Sequenced on the NextSeq 2000 (2X150 cycles), using P4 XLEAP chemistry
• Downsampled to 50M PE reads total across all samples

Phase 2: Operator variability with small panels using the xGen Pan-Cancer, comparing directly against manually preparations

• 20 ng Horizon’s OncoSpan cfDNA prepared using the xGen™ cfDNA & FFPE DNA Library Prep v2 MC Kit, xGen Pan-Cancer Hyb Panel
• This panel has approximately 7800 probes spanning 800 kb of target region for onco-genes
• Sequenced on the NextSeq 2000 (2X150 cycles), using P1 XLEAP chemistry
• Downsampled to 4M PE reads total across all samples

‍Phase 3: Performance of high-plexity libraries on Callisto

• 100 ng HG002 prepared using the xGen™ DNA Lib Prep EZ for exome, xGen Exome Hyb Panel v2
  • 4 captures were performed: 1X, 8X, 12X, 20X
• With higher plexity libraries with 20 pooled samples, you have a much lower ratio of probe:sample
• We are assessing the limits of performance with this kit with various plexities, from 1-plex all the way up to 20-plex
  • With 20-plex, some of the samples are extremely viscous, with 10 µg of DNA in the pool

Key takeaways:

• Walk-away, automated solution for complex end-to-end hybridization capture workflows
• High quality, high-plexity enrichment performance independent of operator
• Consistent coverage uniformity and high on-target bases for different panel sizes
• Callisto reduces hands on time, saving operators 2.5 hours of hands-on time
• Flexible throughput to match lab needs
• Minimal training required with less than 30 minutes of set-up time per run

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Contact our team to discuss how you can automate hybridization capture with the Callisto Sample Prep System.

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