TaqMan™ Mutation Detection Assay with castPCR™ Technology, Gene Reference (75 reactions), Inventoried - FAQs

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).

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.

 

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:

Digital PCR Assays and Kits

QuantStudio Absolute Q Digital PCR System

Absolute Q 1-Step RT-dPCR Master Mix (4X) (Cat. No. A55146) User Guide

TaqMan Mutation Detection Assay with castPCR Technology (Cat. Nos. 4465804, 4465807)

TaqMan Mutation Detection Assay: Competitive Allele-Specific TaqMan PCR

Find additional tips, troubleshooting help, and resources within our TaqMan Primers and Probes Support Center.