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Sequencing is increasingly used in the diagnosis and monitoring of diseases including cancer. With gold-standard accuracy and the ability to detect new genetic variants, Sanger sequencing enables precision medicine.
Kirsten rat sarcoma viral oncogene homologue (KRAS) is a hyper-mutated oncogene across all cancer types. Tumors with KRAS mutations are resistant to many chemotherapeutics including those targeting the epidermal growth factor receptor (EGFR). KRAS mutations can also develop during treatment. Additionally, treatments targeting specific KRAS mutations are currently on the market with more under development.
Ninety-eight percent (98%) of KRAS mutations occur within codons 12 or 13 (exon 2) and codon 61 (exon 3). A large variety of mutations within these codons exist and their frequency varies by cancer type. Sanger sequencing offers several key attributes useful for the detection of KRAS mutations.
Figure 1. Simplified, accurate workflow to detect KRAS mutations. Using reagents such as the BigDye Direct kit saves steps. The protocol is flexible and can be used with a variety of instruments.
Thermo Fisher Scientific has developed a complete and streamlined protocol for the detection of unknown and common KRAS variants including G12C, G12D, and G13D.
We identified primers for the detection of unknown and common KRAS variants including G12C, G12D, and G13D.
Order the same assays referenced in the KRAS variant identification protocol guide by following these steps:
Step 1: Use our Primer Design Tool to search for validated primers using these Assay IDs:
Step 2: Add the primers with the below selections:
The SeqStudio Flex Series genetic analyzers are our most advanced mid-throughput genetic analyzers with the flexibility, usability, connectivity, and serviceability to keep your laboratory up and running at maximum efficiency. Get critical results faster with urgent sample reprioritization capabilities and complete scheduling flexibility.
The BigDye Direct Sanger Sequencing Kit helps enable accurate data. All steps from PCR to capillary electrophoresis are performed in a single tube, reducing hands-on time and the possibility of introducing experimental variability.
For Research Use Only. Not for use in diagnostic procedures.