Catalog number: A35865
The Oncomine Breast cfDNA Assay v2 is part of a complete solution to detect breast tumor-derived DNA (ctDNA) in cell-free DNA (cfDNA). It provides the reagents for library construction and a single pool of multiplex PCR primers for preparation of amplicon libraries from cfDNA obtained from the plasma fraction of a single 10 mL tube of whole blood.
The Oncomine Breast cfDNA Assay v2 enables the analysis of:
Hotspot genes: AKT1, EGFR, ERBB2, ERBB3, ESR1, FBXW7, KRAS, PIK3CA, SF3B1, TP53 (~152 hotspots)
Copy number genes (CNVs): CCND1, ERBB2, FGFR1
Full length genes: TP53 (~80% coverage)

These genes have been identified as frequently mutated in breast cancer. Through the use of Tag Sequencing technology, low limits of detection (LOD) can be achieved for different variant types*:
For SNVs/short indels, an LOD of 0.1% can be achieved with sensitivity of ~80% and specificity of >99%
For detection of TP53 mutations, an LOD of 0.5% can be achieved
For ERBB2 and FGFR1 CNV targets, detection as low as 1.2-fold amplification can be achieved and for CCND1 CNV target, detection as low as 1.4-fold amplification can be achieved with sensitivity of >90% and specificity of >99%
*Sensitivity and specificity for each variant type were determined using a collection of contrived positive samples and cfDNA isolated from normal healthy donors.
Advantages:
Optimized amplicon design for short cfDNA ensures highest possible capture rate
Optimized targeted assay design allows highly multiplexed next-generation sequencing (NGS), reducing sequencing costs per sample
Efficient workflow in just two days

Analysis of SNVs and short indels can be achieved using Torrent Suite Software 5.2 or higher. In order to analyze SNVs, short indels, and CNVs, Ion Reporter 5.6 (cloud- or server-based) is required