B-RAF Mutation Test Kit

Recent studies have shown a correlation between the B-RAF V600E mutation and the development of melanomas. This B-RAF kit will aid current investigations about the link between patient mutation status and response to various cancer therapies. The kit detects the V600E mutation in the B-RAF oncogene in a real-time PCR assay. The mutation is commonly found in melanomas, lung cancer and thyroid cancer.

The methods used in this kit are highly selective and, depending on the total amount of DNA present, can detect approximately 1% of mutant in a background of wild type genomic DNA.  The assays have limits of detection of 5 copies.  These selectivity and detection limits are superior to technologies such as dye terminator sequencing.

 

Intended Use

The DxS B-RAF Mutation Kit is for research use only.


Technological Principles

DxS has combined ARMS™ (allele specific PCR, 6) with the Scorpions™ real-time PCR technology to develop sensitive and robust tests for V600E mutation in the B-RAF oncogene.  
 

Protocol

After DNA extraction, real time PCR assays are performed to detect the target molecule.  By comparing control and mutant sample reactions users can detect and estimate low levels of mutation. No further sample processing is necessary and the time to result is <3 hours
 

Key Features:

  • Provides a qualitative assessment of B-RAF mutation status using DNA samples.
  • Highly selective and can detect less than 1% of mutant in a background of wild type genomic DNA. 
  • The assay has a limit of detection of 5 copies.
  • Simple and easy to follow real-time PCR protocol.
  • Sample types include human genomic DNA from fresh, frozen or paraffin embedded tissue.
     

References

  1. M. Shinozaki, S.J. O’Day, M. Kitago. F. Amersi, C. Kuo, J. Kim, H.J. Wang, D.S.B. Hoon. (2007).  Utility of Circulating B-RAF DNA Mutation in Serum for Monitoring Melanoma Patients Receiving Biochemotherapy.  Clin Cancer Res 13(7): 2068-2074.

  2. N. Dhomen, M. Marais. (2007). New insight into BRAF mutations in cancer.  Current Opinion in Genetics and Development 17: 31-39.

  3. N.E. Thomas. (2006). BRAF somatic mutations in malignant melanoma and melanocytic naevi.  Melanoma Research 16(2): 97-103.

  4. E. Edlundh-Rose, S, Egyhazi, K. Omholt, E. Mansson-Brahme, A. Platz, J. Hansson, J. Lundeberg. (2006). NRAS and BRAF mutations in melanoma tumours in relation to clinical characteristics: a study based on mutation screening by pyrosequencing.  Melanoma Research 16(6): 471-478.

 
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