qBiomarker Somatic Mutations PCR Assays

For detecting the presence of specific DNA sequence mutations in cancer and oncogenesis, these assays from QIAGEN identify individual specific sequence mutations present in cell lines or research samples that are critical for toxicological, drug development, and cancer studies. Mutations are selected from comprehensive curated databases (e.g., COSMIC) and literature reviews based on their clinical or functional relevance and frequency in patient populations.

After isolation of genomic DNA from fresh, frozen, or fixed samples, an aliquot of each sample is added to a separate real-time PCR tube containing the appropriate master mix (included) and the assay, and to a separate tube containing the master mix and the corresponding reference gene copy assay. C_T values are then determined for each mutation-specific assay and reference gene copy assay, and the values are pasted into the appropriate Excel-based data analysis template. Two data analysis methods are supported: the ΔΔCt Method, recommended for experiments using a small (four or less) number of fresh or frozen samples, or a large number of samples with similar DNA quality; and the Average Ct Method, recommended for FFPE samples, large numbers of samples, or samples without wild-type controls. Analysis can be carried out on the somatic mutation data analysis web portal or via downloadable Excel templates.

Designed for routine use on any PCR instrument, the assays are offered in multiple formats compatible with real-time PCR systems from Bio-Rad, ABI, Stratagene, Eppendorf, and Roche LightCycler 480, as well as QIAGEN Rotor-Gene Q/6000 instruments. For FFPE samples, an average Ct below 32 is recommended to allow sensitive detection of mutations; samples that meet this criterion perform robustly on the assays.