PI3K Mutation Test Kit
For sensitive and site-specific detection of 4 somatic mutations in the PIK3CA oncogene in less than 4 hours
The PI3K Mutation Test Kit is a research use only assay for detection of specific mutations in the PIK3CA gene using real-time PCR. The kit provides all necessary reagents optimized for rapid and sensitive detection of a low percentage of mutant DNA in a background of wild-type genomic DNA. ARMS PCR technology combined with the Scorpions detection technology provides high sensitivity and specificity for the detection of specific mutations. Please inquire for more information about the real-time PCR cyclers that are compatible with this kit.
For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.
The PI3K Mutation Test Kit is intended to detect 4 mutations in codons 9 and 20 of the PIK3CA gene. The kit detects the presence of the E545K and E545D mutations but does not discriminate between them.
The kit utilizes two technologies — ARMS (Amplification Refractory Mutation System) and Scorpions — for detection of mutations in real-time PCR.
Allele- or mutation-specific amplification is achieved by ARMS. Taq DNA polymerase is effective at distinguishing between a match and a mismatch at the 3' end of a PCR primer. Specific mutated sequences are selectively amplified, even in samples where the majority of the sequences do not carry the mutation. When the primer is fully matched, the amplification proceeds with full efficiency. When the 3' base is mismatched, only low-level background amplification occurs.
Detection of amplification is performed using Scorpions. Scorpions are bifunctional molecules containing a PCR primer covalently linked to a probe. The fluorophore in this probe interacts with a quencher, also incorporated into the probe, that reduces fluorescence. When the probe binds to the amplicon during PCR, the fluorophore and quencher become separated. This leads to an increase in fluorescence from the reaction tube.
The PI3K Mutation Test Kit includes a control assay and 3 mutation assays. The control and mutation assays can be analyzed in the same PCR run, or they can be used in a two-step procedure. The first step is performance of the control assay to assess the total DNA in a sample. The second step is to complete the mutation assay for the presence or absence of mutated DNA.
The PI3K Mutation Test Kit is intended to detect 4 mutations in codons 9 and 20 of the PIK3CA gene.
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