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
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.
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).
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.