You can now automate your Twist® Bioscience Human Core Exome Kit on the PerkinElmer® Sciclone® NGSx Workstation!
Efficient library construction for Next Generation Sequencing (NGS) of human whole exome sequencing (WES) is fundamentally improved with the superior oligo prob design and performance of the Twist® Human Core Exome Kit. This kit enables increased uniformity and enables increased uniformity and deeper coverage of difficult regions while driving down the cost per sample. When this crucial technology is paired with the PerkinElmer® Sciclone® G3 NGSx workstation, users enjoy interface-guided workflow set-up and advanced step tracking (Figure 1), in addition to the higher throughput and reduced variability afforded by automation.
Metrics generated with automated library preparation, such as yield, mapping rate, coverage, and GC bias are comparable to or superior in performance to manual preparation observed.
Automation protocol for the construction of Twist® Human Core Exome Libraries using the Sciclone® G3 NGSx Liquid Handler
A simplified method section is detailed below. To see the full methods section please see the technical application note.
- Exome libraries (48 pre-capture Combinatorial Dual Index Long Y-Adapter (CDI Standard Protocol) and 48 pre-capture Universal Dual Index Adapter (UDI Protocol)) were generated on a Sciclone® G3 NGSx workstation using Twist® Bioscience Human Core Exome kit with 50 ng NA12878 genomic DNA inputs.
- Samples and libraries were quantified using fluorescence quantification (Thermo Fisher® Qubit® fluorometer)
- Libraries were pooled by mass for subsequent hybridization and target enrichment (8 gDNA libraries per exome hybridization reaction)
- Post capture library pools (2 CDI and 2 UDI) were taken from hybridization through 2 x 76 bp paired-end sequencing on an Illumina® NextSeq® sequencer
- Data was demultiplexed and down sampled to 150X and analyzed using the Picard Metrics Analysis Suit with a minimum Q-score of 20
Analysis of Twist® Human Core Exome Libraries Generated on the Sciclone G3 NGSx liquid handler
The NGS libraries generated from PerkinElmer’s® Sciclone® G3 NGSx workstation protocols successfully built libraries with yields (pre-capture concentration approximately 80 ng/µL, post-capture concentration approximately 10-20 ng/ µL) and sizes (pre-capture 343 bp, post-capture 346.3 bp) within the expected range of the Twist® Human Core Exome manual (Table 1).
The sequencing of the exome targeted enrichment libraries was compared for metrics such as: coverage, GC percentage and mapping quality (Table 2).
Simplify the Twist® Human Core Exome Library Prep Workflow while Generating High Quality Libraries
Overall, the libraries concentration, mean size, and sequencing statistics indicate that automating the Twist® Bioscience Human Core Exome Kit workflow on the PerkinElmer® Sciclone® G3 NGSx workstation generated libraries of similar or superior quality when compared to manual preparation. This combination of products provides a highly simplified workflow for generated high quality exome sequencing libraries with user-friendly automation capabilities.
Listen to Dr. Renata Pellegrino talk about how she scaled up her Twist® Target Capture workflow here.