QC validation by NGS guarantees uniformity of coverage which combined with error suppression technology ensures confident calling of all mutations down to 0.1% VAF.

Our online probe design tool means you can design and optimise a panel in minutes and our rapid manufacture process means you will receive your panel within 4 weeks of ordering.

Cell3™ Target enrichment has been optimised to deliver efficient target capture and unrivalled on target performance regardless of panel size from 1 gene to 1,000.

While detection of low frequency mutations <20% VAF has been possible with NGS methods for a number of years, accurate detection of ultra-low frequency mutations (below 5% VAF) has been hampered by excessive false positives or artefactual variant calls.

The need to accurately call variants below 5% VAF is important for the advancement of non-invasive techniques like liquid biopsy testing where the sample used, circulating tumor DNA (ctDNA), is in low abundance and requires ultra-sensitive sequencing methods to detect it.

Targeted sequencing through the use of custom designed NGS panels offers a cost-effective method for rare variant detection. Without the data burden of whole-genome sequencing (WGS) or whole-exome sequencing (WES) it allows researchers to achieve very high depths of coverage providing the increased sensitivity and accuracy needed for ctDNA analysis. However, deep sequencing is not without its flaws as sequencing low-abundance DNA at high depth also increases the level of sequencing artefacts, creating background noise and potentially masking low-frequency variants. In light of this, it is essential when you are working with ctDNA that extra care is taken to ensure that errors are suppressed, and only high-quality variant calls are retained.

Cell3™ Target is a novel target enrichment system developed by Nonacus for converting any type of DNA (cfDNA, gDNA, FFPE DNA) into libraries for next generation sequencing. It uses error suppression technology to ensure confident calling of all mutations down to 0.1% VAF and is ideal for rare variant detection in liquid or tissue biopsies.

We understand that our customers have different questions to answer so Cell3™ Target has been developed for and validated on ctDNA, gDNA and FFPE DNA for a range of oncology applications, so that whether you are investigating early cancer detection, diagnosis and treatment selection, tumor heterogeneity or using ctDNA for tumor monitoring purposes, you know you will be able to use the same method and panel design.

Cell3™ Target enrichments incorporate error suppression technology including unique molecular indexes (UMIs) and unique dual indexes (UDI’s) which remove of both PCR and sequencing errors and index hopping events. This error suppression technique, combined with our excellent uniformity of coverage, allows you to confidently and accurately call variants down to 0.1% VAF and enables generation of sequencing libraries from as little as 1 ng of cfDNA input. We provide advice and provision of ready to go analysis scripts for error removal using UMI’s.

Figure 1. Using UMI’s to identify and quantify individual DNA molecules during library preparation increases sensitivity

Good design is key to a successful NGS panel ensuring adequate coverage and robust variant calling.  It’s not always easy to achieve, especially first time, but our Probe Design tool, makes it simple and easy allowing you to optimise panels in minutes. Create designs instantly from scratch using uploaded BED files, gene lists or genomic coordinates, customise catalogue products or add your own content onto one of our fixed panels.

• Easily merge, compare and share panels

• Get coverage data instantly and request a quote

• Receive your fully validated* custom panel within 4 weeks of order.

Log into the Probe Design Tool at MyNonacus  to start on a design and get a quote.

*All panels are NGS validated to ensure uniformity of coverage meets our QC requirements across all targeted regions

The baits used in Cell3™ Target panels are designed to deliver excellent uniformity of coverage. By improving uniformity of coverage and reducing the number of low coverage exons, our targeted enrichment optimises sequencing efficiency and sample capacity per sequencing run ultimately reducing your sequencing costs.