How can I find a mutant control sample for my Mutation Detection experiment?
You can check the COSMIC database to see if a control sample is identified for your mutation. If one is not available, you can have one synthesized (using our GeneArt Gene Synthesis service, for example).
In a Mutation Detection experiment, what is the difference between the wild-type assay and the gene reference assay?
The wild-type assay targets the same site as the mutant assay, but blocks the mutant allele. The reference assay detects the gene of interest at a mutation-free region on the same gene. The gene reference assay will amplify both mutant and wild-type gDNA, while the wild-type assay will only amplify the wild type. Both the reference assay and the wild-type allele assay can be used in the same manner to determine the percent mutation status of a sample. Wild-type allele assays are only available for the initial subset of assays.
Can I use TaqMan SNP Genotyping Assays to detect single-nucleotide variants (SNVs) that are present in less than 5% of a sample?
TaqMan SNP Genotyping Assays are designed to work on equal volumes, so if the mutation is only present at 5%, it is possible that the assay will not detect the variants. Therefore, we do not recommend using these assays to detect somatic mutations. SNP assays by qPCR with TaqMan SNP Genotyping Assays are intended to detect germline mutations.
For somatic mutations, we recommend using digital PCR or castPCR. Find more information about digital PCR and castPCR in the links below: