Automate Complex Next Generation Sequencing Library Construction!

The Sciclone® G3 NGSx iQ workstation (Figure 1) is specifically designed to automate the most complex Next Generation Sequencing (NGS) library construction workflows. The workstation utilizes:

  • On-deck thermal cycler (ODTC) capability
  • Integrated tip storage with robotic tip transfer
  • Highly reproducible liquid transfer technology
  • On-deck thermal elements
  • The Twister® III robot arm

This technology enables complex processes such as:

  • Automated cDNA synthesis in RNA-seq workflows
  • Polymerase Chain Reaction (PCR)
  • Fragmentation steps which occur at high temperatures
  • Adenylation and end repair

Figure 1: The Sciclone® G3 NGSx iQ workstation offers integrated tip storage, an on-deck thermal cycler, and highly reproducible liquid transfer technology which reliably constructs high-quality NGS libraries from DNA or RNA.

Roche® KAPA® HyperPlus kit with Roche® KAPA® Adapters and Roche® KAPA® Pure Beads is a complete library preparation solution compatible with Illumina® Sequencing platforms. The workflow (Figure 2) allows higher throughput among others, seen in our technical note.

Figure 2: With only thirty minutes of manual bench setup required, a researcher can walk-away and focus on intellectual contributions such as data analysis or experimental design.

Automation Protocol for the Construction of DNA Libraries with Roche® KAPA® HyperPlus Kit

Libraries were prepared according to the following protocol:

  • 96 libraries were prepared on the Sciclone® G3 NGSx iQ workstation
  • 100 ng control DNA NA12878 (Coriell Institute) was used with the Roche® KAPA® HyperPlus Kit
  • 3 cycles of PCR amplification followed

Following completion of the automated library preparation, the Thermo Fisher® Scientific Qubit® HS assay was used for quantification, and peak size distribution was determined by the LabChip® GX Touch nucleic acid analyzer with the NGS 3K assay (Figure 3).

Figure 3: LabChip® GX Touch nucleic acid analysis of 12 libraries generated across a 96-well PCR plate from the Roche® KAPA® HyperPlus kit.

Yields and Sizes of Automated Libraries were Equivalent to Manually Prepared Libraries

Overall the yields (29.1 ± 4.8 ng/μl) and sizes (415 ± 74.7 base pairs) of the NGS libraries generated with the Sciclone® G3 NGSx iQ workstation were within the expected range of libraries prepared manually using the Roche® KAPA® HyperPlus kit and Roche® KAPA® Index barcodes.

Roche® KAPA® HyperPlus Automation on the Sciclone® G3 NGSx iQ™ Workstation Increases Throughput and Reproducibility of High-Quality NGS Libraries

The Roche® KAPA® HyperPlus kit automated on the Sciclone® G3 NGSx iQ™ workstation delivers uniform NGS libraries with yields and sizes consistent with manually prepared libraries and enables complete walk-away library preparation which facilitates error minimization, reduces hands-on time, and increases throughput and reproducibility.

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For research use only. Not for use in diagnostic procedures.