The Swift Normalase Kit is a revolutionary, enzymatic library normalisation technology for next-generation sequencing (NGS) from Swift Biosciences. Developed to save time and increase throughput by providing uniform sample processing with fewer handling steps, the Swift Normalase Kit is compatible with diverse library preparation methods and sample types to produce more evenly balanced sequence data.
Advantages of Normalase over traditional library quantification methods
• Save time and increases throughput
• Reduce variability to save on sequencing costs
• Flexible design for many workflows
The Swift Normalase workflow
Following conventional amplification with Normalase primers to generate libraries with indexed adapters to ≥12nM, a standard purification is performed. 4nM of each library is then enzymatically selected with Normalase I and the libraries are combined into a single pool. Next, the Normalase II enzymatically normalises all libraries within the pool to 4nM and thus, without any need for further purification, the pool is ready for sequencing:
Libraries supported by Swift Normalase include those with full-length indexed adapters (e.g. Accel-NGS 2S, Swift 2S Turbo, KAPA®, Illumina®, TruSeq®), and other libraries with an amplified yield of ≥12nM, or which have been prepared for direct sequencing (e.g. whole genome, whole transcriptome). By processing these with Swift Normalase, researchers can benefit from reduced variability, higher multiplexing per run and fewer sample failures.
Amplification with Normalase primers is validated with multiple polymerases
A. Libraries generated with 50ng Coriell NA12878 DNA were amplified for 6 cycles with Normalase primers using three different DNA polymerases; NEB Q5 HiFi, Kapa® HiFi, and Swift HiFi (n=5 libraries/polymerase). All polymerases tested amplified libraries to ≥12nM, meeting the minimum threshold for Normalase. B. Amplified libraries were normalized with Normalase and sequenced on the Illumina® MiSeq using a V2 micro flowcell 1x50 cycles. Performance of Normalase normalization was equivalent across the three HiFi polymerases. The variation across libraries was calculated at a CV of 8.7%.
Material available for download
Normalase - a novel library normalisation tool for high-throughput NGS
Webinar: Radically streamlining NGS library quantification and normalisation
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KAPA® is a registered trademark of ROCHE MOLECULAR SYSTEMS, INC.
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