LightMix® Digital BRAF V600E
The LightMix® Digital BRAF V600E allows the detection of the V600E mutation in gene BRAF (COSMIC ID: COSM476) in genomic human DNA from a nucleic acid extract. The point mutation V600E substitutes valine by glutamic acid at amino acid position 600 of the protein serine/threonine-protein kinase B-Raf. This mutation can activate the protein function.
Using PCR methodology, a 117 bp fragment covering the BRAF V600E mutation is amplified with specific primers. The mutation site in the PCR fragment is analyzed using two labeled probes binding competitively to the mutation site. The mutant probe is FAM- labeled, and the wild type (WT) probe is HEX-labeled. The PC included in the kit contains 4-6 % synthetic mutant target in wild-type genomic DNA and is equivalent to ~17 ng/µl when dissolved in 60 µl of PCR grade water. It is recommended to always include at least one PC and a No-Template-Control (NTC) (e.g. PCR grade water) in each run.
