Includes all controls.

BRAF Mutations and Cancer

The BRAF gene encodes a serine/threonine protein kinase, which plays a role in regulating the MAP Kinase/ERK signaling pathways, affecting cell growth and proliferation. Somatic mutations in this gene have been found in cancers, including non-Hodgkin lymphoma, colorectal cancer, malignant melanoma, papillary thyroid carcinoma, non-small cell lung carcinoma, and adenocarcinoma of lung.The most common mutations in BRAF occur in codon 600, where an amino acid substitution in the activation segment of the kinase domain creates a constitutively active form of the protein. BRAF mutations are generally found in tumors that are wild-type for KRAS, NRAS, and EGFR.

Testing Procedure and Analysis

The table below lists the mutations detected by this kit and the approximate frequencies found in various cancers.

Mutation Name Nucleotide Change Amino Acid Change Frequency
V600E c.1799T>A Valine (V) to glutamic acid (E) 70-90%
V600K c.1798_1799GT>AA Valine (V) to lysine acid (K) 10-15%
V600D c.1799_1800TG>AT Valine (V) to aspartic acid (D) <5%
V600R c.1798_1799GT>AG Valine (V) to arginine (R) <5%
V600M c.1798G>A Valine (V) to methionine (M) <1%
V600G c.1799T>G Valine (V) to Glycine(G) <1%

EntroGen’s BRAF codon 600 mutation analysis kit is a polymerase chain reaction (PCR)-based assay that uses allele-specific probes to identify the presence of the BRAF V600E, V600K, and V600D mutations (V600R and V600M included in the BRAFX-RT64 kit). The testing procedure involves three (3) simple steps:

  • Isolation of DNA from tumor biopsies, paraffin-embedded sections (FFPE), or fresh frozen tumors
  • Amplification of regions of the BRAF gene using allele-specific probes.
  • Detection of amplification product on a Real-Time PCR instrument.

This test can be completed in approximately 2 hours from isolation of DNA to test result.

Equipment and Materials

EntroGen’s BRAF codon 600 mutation analysis kit requires a real-time PCR instrument capable of detecting FAM and VIC fluorescent probes. This test includes reagents required for the PCR amplification/detection, as well as validated reaction controls. Columns and reagents for DNA isolation are not included.

Intended Use

USA: EntroGen’s BRAF Codon 600 mutation analysis kit is provided for research use only (RUO). Not for use in diagnostic procedures.

Europe: EntroGen’s BRAF Codon 600 mutation analysis kits (BRAFM-RT64 and BRAFX-RT64) are available for research (RUO) and diagnostic (CE-IVD) purposes.

Bibliography of studies using this product

Kimbrell HZ, Sholl AB, Ratnayaka S, Japa S, Lacey M, Carpio G, Bhatia P, Kandil E. BRAF Testing in Multifocal Papillary Thyroid Carcinoma. Biomed Res Int.  2015;2015:486391. Pubmed ID 26448939

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