The KRAS Mutation Analysis Reagents employ a simple fragment analysis workflow to detect 1% mutant to wild type genomic DNA across 12 common and rare mutations in the KRAS gene region (see Table 1). The KRAS reagent set includes everything needed for the analysis of 30 samples.

Advantages of the KRAS Analysis Reagents include:
• Simple Analysis—unambiguous, easy-to-interpret results
• Thorough Coverage— detects 12 mutations in the KRAS 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

KRAS Mutation Detection Plays 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 conditions (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 information about our BRAF Mutation Reagents

Keep Pace with Current EGFR Therapy Research
The Applied Biosystems® KRAS Mutation Analysis Reagents were developed to facilitate research aimed at elucidating the role of this regulatory protein in Oncology. Offering an off-the-shelf solution, the KRAS reagent set provides a straightforward protocol with results analyzed on the industry-standard 3500 Series or 3130 Series Genetic Analyzers. This allows you to easily add KRAS mutation analysis to your laboratory´s genotyping capabilities.

Simple, Streamlined Workflow for Accurate KRAS Mutation Analysis
The protocol for the KRAS Mutation Analysis Reagents is simple and efficient (Figure 2). 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 (Table 1).

Additionally, this reagent set is optimized using the same PCR thermal cycling conditions and run modules for capillary electrophoresis that are used for analysis of the BRAF gene on either the 3500 Series or the 3130 Series Genetic Analyzers. View information about our BRAF Mutation Reagents

Shifted Termination Assay (STA) Yields Unambiguous Results
KRAS 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 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, followed by data analysis 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.

References
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.