TaqMan Assays

The use of TaqMan® MGB probes, developed by Applied Biosystems (ABI), for the detection of single nucleotide polymorphisms (SNPs) allow us to increase the specificity of PCR assays. The principle of the TaqMan probe relies on the 5’-3’ nuclease activity of the enzyme Taq-polymerase and fluorophore-based detection:

TaqMan® assays contain two specific primers targeting the region flanking the SNP site and two TaqMan fluorescent probes with a Minor Groove Binder (MGB).

Each probe is labeled with a different fluorophore (VIC or 6-FAM). These reporters are attached covalently to the 5' end of the two probes. Near the 3 'end, there is a non-fluorescent quencher (NFQ) that prevents  liberation of the reporter fluorescence if the probe is not degraded. The MGB serves to stabilize the double-stranded structure formed between the target and the probe.

During the PCR reaction, probes that hybridize specifically to DNA fragments are destroyed and the fluorescence of corresponding fluorophore is liberated.

Shematic depiction of a TaqMan Genotyping Assay (source: ABI)

Applications

SNP Genotyping

TaqMan® SNP Genotyping Assays make it easy to perform SNP genotyping studies with the precision of TaqMan® chemistry. TaqMan® assays provide SNP detection capabilities with high-confidence results.

DNA copy number analysis

TaqMan® Copy Number Assays are designed for copy number analysis of copy number variations (CNVs) and smaller genomic regions. CNV is a polymorphism associated with diseases such as cancer, immune diseases and neurological disorders as well as drug metabolism.

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