The BRAF Mutation Analysis Reagents employ a simple fragment analysis workflow to detect 1% mutant to wild type genomic DNA across 3 common and rare mutations in the BRAF gene region (see Table 1). The BRAF set provides sufficient reagent for analysis of 30 samples.
Advantages of the BRAF Mutation Analysis Reagents include:
• Simple Analysis—unambiguous, easy-to-interpret results
• Thorough Coverage—detects 3 mutations in the BRAF gene
• Sensitive—detects mutations at frequencies as low as 1 to 5% against a wild type genomic DNA background
• Efficient—interrogate multiple loci from the same sample in a minimum number of tubes
BRAF Mutation Detection Play Vital Role in mCRC Research
The epidermal growth factor receptor (EGFR) pathway is a complex signaling cascade that preliminary research has associated with the development and progression of many cancer types (Figure 1). KRAS and BRAF gene mutations are present in a number of cancers including those of the colon, lung, pancreas, biliary tract, endometrium, and ovary. It has been shown that approximately 35% to 45% of metastatic colorectal cancer (mCRC) tumors may have a KRAS or BRAF mutation [1,3], which makes them less likely to respond to anti-EGFR therapies. So identifying these mutations is of great importance in clinical and pharmaceutical research.
View info about our KRAS Mutation Reagents
Keep Pace with Current EGFR Therapy Research
The Applied Biosystems® BRAF Mutation Analysis Reagents were developed to facilitate research aimed at elucidating the role of these regulatory proteins in Oncology. Offering an off-the-shelf solution, the BRAF reagent sets provide a straightforward protocol with results analyzed on the industry-standard 3500 Series or 3130 Series Genetic Analyzers. This allows you to easily add BRAF mutation analysis to your laboratory’s genotyping capabilities.
Simple, Streamlined Workflow for Accurate BRAF Mutation Analysis
The BRAF reagent set analyzes 3 important SNPs in a single multiplex PCR (Table 1), with a workflow that is simple and efficient. For additional information on the workflow (Figure 2) please see the protocol on the Literature⁄Support tab.
Additionally, this reagent set is optimized using the same PCR thermal cycling conditions and the same run modules for capillary electrophoresis on either the 3500 Series or the 3130 Series Genetic Analyzers. View information about our KRAS Mutation Reagents
Shifted Termination Assay (STA) Yields Unambiguous Results
The BRAF Mutation Analysis Reagents employ a proprietary shifted termination assay (STA) technology to amplify the mutation signal. The assay is performed by using specially designed primers, enzyme master mix, and chemistry protocol. The STA reaction recognizes wild-type or mutant target sequences and selectively extends the detection primer with 1 to 20 nucleotides to generate various lengths of primer extension product. The extended STA fragments are separated by capillary electrophoresis with an Applied Biosystems® 3500 Series or 3130 Series Genetic Analyzer, and analyzed using GeneMapper® Software.
GeneMapper® Software v4.1 Offers Easy Data Analysis
GeneMapper® Software is a flexible genotyping software package that provides DNA sizing and quality allele calls for all Applied Biosystems electrophoresis-based genotyping systems. GeneMapper® Software v4.1 offers a simple, qualitative method for interpreting fragment analysis data, where anyone can easily identify mutations present in the sample (Figure 4, 5). To streamline the data analysis, we have developed a bin setup protocol for assistance in BRAF analysis using GeneMapper® Software v4.1 which can be found here.
1. Di Nicolantonio F, Martini M, Molinari, F et al. (2008) Wild-type BRAF is required for response to panitumumab or cetuximab in metastatic colorectal cancer. J Clin Oncol 26(35):5705–5712.
2. Karapetis CS, Khambata-Ford S, Jonker DJ, et al (2008) K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med 359(17):1757–1765.
3. Goncalves A, Esteyries S, Taylor-Smedra B, et al. (2008) A polymorphism of EGFR extracellular domain is associated with progression free-survival in metastatic colorectal cancer patients receiving cetuximab-based treatment. BMC Cancer 8:169.