Reagents for KRAS and BRAF mutation analysis
Designed to facilitate clear, comprehensive, sensitive testing
The Applied Biosystems® KRAS Mutation Analysis Reagents and 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, these KRAS and BRAF reagent sets provide a straightforward protocol with results analyzed on the industry-standard Applied Biosystems® 3500 Series or 3130 Series Genetic Analyzers. Seamlessly integrate KRAS and BRAF mutation analysis into your laboratory’s capabilities and maximize the utility of your Applied Biosystems® genetic analysis instrument.
- Simple analysis — clear, easy-to-read results
- Comprehensive coverage — detects 12 mutations in the KRAS gene; 3 mutations in the BRAF gene
- Sensitive — able to detect 1–5% mutation contribution in a background of wild type genomic DNA from analytical samples
- Efficient — interrogate multiple loci from the same sample, using minimal numbers of test tubes
The epidermal growth factor receptor (EGFR) pathway is a complex signaling cascade that is associated with the development and progression of many cancer conditions. 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, which makes them less likely to respond to anti-EGFR therapies. Identifying these mutations is greatly important to clinical and pharmaceutical research.
The KRAS and BRAF Mutation Analysis Reagents employ a proprietary shifted termination assay (STA) technology to amplify the mutation signal. The assay is performed 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 products. The extended STA fragments are separated by capillary electrophoresis with an Applied Biosystems® 3500 Series or 3130 Series Genetic Analyzer, followed by data analysis using GeneMapper® Software.
The protein products of the KRAS and BRAF genes perform essential functions in the epidermal growth factor (EGF) signaling pathway. Because of its involvement in the regulation of cell proliferation, the EGF pathway is studied by many cancer researchers.
|Mutations Interrogated Using the KRAS Mutation Analysis Reagents |
Mutations Interrogated Using the BRAF Mutation Analysis Reagents
|KRAS Codon 12 Mutations||KRAS Codon 13 Mutations||BRAF Mutations|
|Gly12Ser (GGT>AGT)||Gly13Ser (GGC>AGC)||Val600Glu (GTG>GAG)|
|Gly12Arg (GGT>CGT)||Gly13Arg (GGC>CGC)||Val600Gly (GTG>GCG)|
|Gly12Cys (GGT>TGT)||Gly13Cys (GGC>TGC)||Val600Ala (GTG>GGG)|
|Gly12Asp (GGT>GAT)||Gly13Asp (GGC>GAC)|
|Gly12Ala (GGT>GCT)||Gly13Ala (GGC>GCC)|
|Gly12Val (GGT>GTT)||Gly13Val (GGC>GTC)|
Simple, Streamlined Workflow for Accurate KRAS and BRAF Mutation Analysis
The protocol for the KRAS and BRAF Mutation Analysis Reagents is simple and efficient . The KRAS reagent set amplifies 12 important SNPs in two multiplex PCR analyses: 6 SNPs for codon 12 in a single multiplex, and 6 SNPs for codon 13 in an additional multiplex. Similarly, the BRAF reagent set analyzes 3 important SNPs in a single multiplex PCR (Table below). Both reagent sets are 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. This allows you to analyze samples amplified by either the KRAS or BRAF reagent sets concurrently and reliably.