The TRUPCR® BRAF Mutation Kit is an in vitro diagnostic test intended for the qualitative detection of BRAF somatic mutations in the genomic DNA extracted from fresh, frozen or formalin fixed paraffin-embedded (FFPE) tissue. The TRUPCR® BRAF mutation kit is based on allele specific amplification and is achieved by ARMS PCR. Taq DNA polymerase is extremely effective at distinguishing between a match and a mismatch at the 3’-end of a PCR primer. The kit is designed to selectively amplify mutant specific sequences in samples that contain a mixture of wild-type and mutated DNA. The most common mutations are found in codon 600. The detection is achieved using fluorescent probes labelled with FAM and HEX. The TRUPCR® BRAF Kit is composed of 7 assays for the detection of the BRAF mutations and a reference control gene of BRAF region without any known polymorphism/mutation.
Key Features:
-
- Selective Amplification of DNA containing mutation with ARMS Technology.
- Sensitive to detect up to 1% mutation in BRAF gene.
- Detects 7 different mutations in a single run.
- Extraction control included to avoid false-negative results.
- Rapid, more reliable, comprehensive and cost effective tests.
- Easy work flow & compatible with various Real Time PCR instruments.
BRAF Detectable mutations:
| TUBE | MUTATION | NUCLEOTIDE CHANGE | COSMIC ID |
|---|---|---|---|
| Tube 1 | V600E | c.1799T>A | COSM476 |
| Tube 2 | V600E Complex | c.1799_1800TG>AA | COSM475 |
| Tube 3 | V600D | 1799_1800TG>AT | COSM477 |
| Tube 4 | V600G | c.1799T>G | COSM6137 |
| Tube 5 | V600K | c.1798_1799GT>AA | COSM473 |
| Tube 6 | V600M | BRAF c.1798G>A | COSM1130 |
| Tube 7 | V600R | c.1798_1799GT>AG | COSM474 |
The RAS/RAF/MEK/ERK pathway is a critical proliferation pathway in many human cancers. This pathway can be constitutively activated by alterations in specific proteins, including BRAF, which phosphorylates MEK on 2 regulatory serine residues. Over 45 cancer-associated mutations have been identified in BRAF. BRAF mutations have been identified at a high frequency in specific cancers, including approximately 50 to 60% of melanoma, Approximately 90% of all identified BRAF mutations that occur in human cancer are a T1799A transversion mutation in exon 15, which results in a V600E amino acid substitution. This mutation appears to mimic regulatory phosphorylation, locks the BRAF kinase in its active status, and increases BRAF activity approximately 10-fold compared to wild type (2). T1799A alteration (V600E mutation) accounts for 70 to 90% of BRAF mutant melanoma patients. In addition, the T1799A alteration could be associated with a second nucleotide mutation (G1798A) and leads to a V600K mutation in an additional ~6% to 29% of patients with a BRAF mutation
Ordering Information:
| CAT. NO. | PRODUCT | CONTENTS |
|---|---|---|
| 3B1287 | TRUPCR® BRAF Mutation Kit | 24 Reactions |
| 3B1288 | TRUPCR® BRAF Mutation Kit | 48 Reactions |