Digital detection of single nucleotide variants (SNVs) and small InDels from as little as 5 ng of DNA from FFPE in a single tube. Designed for compatibility with other Vantage 3D™ Assays, the SNV Assays may be combined with mRNA, gene fusion, and protein for in-depth 3D Biology™ characterization of key oncogenesis pathways.
- Simple protocol, optimized for challenging FFPE samples
- Validated workflows for cell suspensions, fresh/frozen tissue, and FFPE
- Concordance with gold standard mutational analysis techniques
- Sensitivity and Specificity above 95% on solid tumor samples with >5% allele frequency*
- Simple, integrated data analysis, eliminating the need for a bioinformatician
*Validated on internal reference materials. Data may vary with the various quality of FFPE samples.
The nCounter® Vantage 3D DNA SNV Assays leverage NanoString’s proven molecular barcode technology. Each SNV panel is designed to detect the mutant allele of interest and a reference allele using a set of probes with novel architecture. The upstream probe (Probe S) has proprietary sequences that become unstable in the presence of a single nucleotide mismatch. The downstream probe (Probe T) is based on existing NanoString probe chemistry. Pools of Probe S and T are combined with an nCounter® molecular barcode SNV TagSet to enable highly specific detection of SNVs in a background of abundant normal tissue alleles.
Integration with the nSolver™ Analysis Software provides a comprehensive detection solution from sample to data. Due to digital barcode chemistry, variants are detected at 1–10% levels.
In the data shown, all assayed variants are detected with >95% confidence (p<0.05 confidence threshold indicated by the dashed pink line).
In collaboration with a team from The University of Texas MD Anderson Cancer Research Center, 5 ng of purified genomic DNA from over 40 different FFPE clinical research samples from a variety of cancer types were assayed with the nCounter® SNV detection chemistry. DNA integrity scores (DIN from TapeStation 2200 analysis) ranged from 2.3 to 6.1 for these samples. Somatic variants that had been previously detected by next-generation sequencing were detected with 98.1% sensitivity, 100.0% specificity, and 99.9% accuracy. Representative results for a selection of these samples are shown in Table 1.
- Multiplex detection of driver mutations in lung cancer: Simultaneous assay of single nucleotide variants (SNV) and fusion transcripts from small amounts of FFPE samples on the nCounter® analysis system
- Enzyme-free, Amplification-free, Hybridization-based Single Molecule Sequencing (Hyb & SeqTM)
- Oncology Biomarker Development: Simultaneous Digital Counting of Nucleic Acids and Proteins at 800-plex
- Multiplex profiling of cancer driver mutations: Detection of single nucleotide variants and small InDels from small amounts of FFPE samples on the nCounter® analysis system
For Research Use Only. Not for use in diagnostic procedures.