LightMix® Digital PIK3CA E545K
The point mutation E545K substitutes glutamic acid by lysine at amino acid position 545 of the protein phosphoinositide 3-kinase alpha. The protein is a lipid kinase and is involved in fundamental cellular processes such as cell proliferation and differentiation. This mutation has been described to activate constitutively the protein function, which can lead to oncogenesis.
Using PCR methodology, a 83 bp fragment covering the PIK3CA E545K 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.
