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LightMix® Digital PIK3CA E545K

Material Number: 10068507001

The LightMix® Digital PIK3CA E545K allows the detection of the E545K mutation in gene PIK3CA in genomic human DNA from a nucleic acid extract. It is a kit with reagents for a maximum of 32 (HiSense plate) or 46 (Universal plate) digital PCR reactions

Quantity: For a maximum of 32 (HiSense plate) or 46 (Universal plate) digital PCR reactions

Additional information

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.

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