This is the amplicon context sequence in accordance with the minimum information for the publication of real-time quantitative PCR experiements (MIQE) guidelines. For more details, please refer to the following publication, "Primer Sequence Disclosure: A Clarification of the MIQE Guidelines."
ddPCR™ probe assay designed for mutation detection. Probe assays consist of unlabeled PCR primers and a dual labeled fluorescent probe.
Info: 64nt amplicon; crosses exon-intron junction
The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1 which then phosphorylates and inactivates components of the apoptotic machinery. Mutations in this gene have been associated with the Proteus syndrome. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq Jul 2011]
Products used to generate validation data:
Dilution series of mutant DNA in ~40,000 copies of wild type DNA background.
ddPCR Amplitude Scatter Plot (wild type + mutant)
Single-well data for mutant DNA spiked into wild-type DNA (~0.1%). Mutation assay (FAM, Channel 1) was duplexed with the corresponding wild-type reference assay (HEX, Channel 2).
ddPCR Amplitude Scatter Plot (wild type)
Single-well data for wild-type DNA (no mutant). Mutation assay (FAM, Channel 1) was duplexed with the corresponding wild-type reference assay (HEX, Channel 2).