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Sensitive mutation detection for cancer research
Rare mutation detection has great implications in areas such as cancer research because the accumulation of mutations in crucial regulatory genes, such as oncogenes or tumor suppressor genes, is an important aspect of tumorigenesis. Acquisition of these mutations in a tiny subset of somatic cells can be sufficient for cancer initiation or progression.
Since these mutations are so rare, they require an assay that delivers high signal-to-noise and low false-positive-to-false-negative rates.
Common SNP genotyping technologies, such as capillary electrophoresis (CE) sequencing and real-time PCR, are most effective at detecting mutant cells with a prevalence no lower than about 20% (or approximately 1 in 5 cells). By combining real-time PCR chemistries, such as TaqMan® Assays, with digital PCR methodology, researchers are now able to detect mutant cell prevalence down to 1%—and below.
Digital PCR works by partitioning a sample into many individual reactions prior to amplification, reducing competing wild type sequences in any reaction containing a mutation and effectively decreasing background noise. If sufficient partitions are used, the reaction wells reach a point where the wild-type signal no longer overwhelms the mutant signal. Because each data point is generated digitally, the total count of each allele, mutant and wild type, can be calculated and a ratio determined.
Digital PCR: Rare Mutation Applications on the QuantStudio™ 3D
Genetic mutation is a hallmark of cancer, and the ability to detect genomic DNA changes is paramount to understanding this disease at a molecular level. Assays developed to detect rare mutations need to be able to identify just a few mutant cells in a huge abundance of wild type cells. Digital PCR combined with TaqMan SNP Genotyping Assays offer a highly sensitive and specific solution for the confirmation of these discoveries.
Poster: Utilizing the QuantStudio 3D Digital PCR System for BRAF V600E Mutation Detection in Papillary Thyroid Carinoma and Malignant Melanoma
Product Bulletin: QuantStudio™ 3D Digital PCR System
User Guide: QuantStudio™ 3D Digital PCR System
Digital PCR Experiment Design Guide
QuantStudio™ 3D Digital PCR System
Absolute quantification, using sealed-chip technology, for a reliable method and precise, sensitive data.
For Research Use Only. Not for use in diagnostic procedures.