Taqman 5 nuclease

Manufactured by Thermo Fisher Scientific

Sourced in United States

The TaqMan 5' nuclease assay is a real-time PCR technology used for the detection and quantification of specific DNA sequences. It utilizes a fluorogenic probe that binds to the target DNA sequence, and the 5' nuclease activity of Taq polymerase cleaves the probe, resulting in the release of a fluorescent signal that is proportional to the amount of target DNA present in the sample.

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Lab products found in correlation

Massarray iplex, by Labcorp (1 mentions) Human omni express exome bead chip kit, by Illumina (1 mentions)

3 protocols using taqman 5 nuclease

Genotyping of Diabetes SNPs in MDCS

Cited 1 time

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A total of 58 previously reported type 2 diabetes-associated single nucleotide polymorphisms were selected for genotyping in the MDCS (Additional file 1: Table S1) [24 (link), 28 (link)]. Genotyping was performed by MassARRAY iPLEX (Sequenom, San Diego, CA, USA) or Taqman 5′ nuclease (Applied Biosystems, Foster City, CA, USA) assays according to the manufacturers’ instructions. The concordance rate was >99 % in 5500 samples which were genotyped using Human Omni Express Exome Bead Chip Kit (Illumina, San Diego, CA, USA). Genotyping success rate ranged between 96 and 99 %. All genotypes were in the Hardy-Weinberg equilibrium (P > 0.0009).

Hindy G., Mollet I.G., Rukh G., Ericson U, & Orho-Melander M. (2016). Several type 2 diabetes-associated variants in genes annotated to WNT signaling interact with dietary fiber in relation to incidence of type 2 diabetes. Genes & Nutrition, 11, 6.

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Quantitative RT-PCR Gene Expression

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TaqMan 5’ nuclease quantitative (real-time) RT-PCR assays were carried out using pre-developed assays from Applied Biosystems according to the manufacturer’s protocol. Reactions were carried out in triplicate. The endogenous controls GUSB and/or PP1A were tested in separate reactions. Relative expression levels were assessed by the comparative threshold cycle (CT) method.

Abdel-Rahman M.H., Sample K.M., Pilarski R., Walsh T., Grosel T., Kinnamon D., Boru G., Massengill J.B., Schoenfield L., Kelly B., Gordon D., Johansson P., DeBenedictis M.J., Singh A., Casadei S., Davidorf F.H., White P., Stacey A.W., Scarth J., Fewings E., Tischkowitz M., King M.C., Hayward N.K, & Cebulla C.M. (2019). Whole exome sequencing identifies candidate genes associated with hereditary predisposition to uveal melanoma. Ophthalmology, 127(5), 668-678.

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Genetic Profiling of Inflammatory Cytokines

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The following locations were investigated in this study: IL-1β –31 T/C (rs1143643), IL-6 –174 G/C (rs1800795), TNF-α –308 G/A (rs1800629), and IFN-γ +874 A/T (rs2430561) genes. Genotyping was performed with TaqMan^® 5’ nuclease allelic discrimination (Assay by Design/Demand, Applied Biosystems, Foster City, CA) as described previously [43 (link)–45 (link)].

Allam G., Alsulaimani A.A., Alzaharani A.K, & Nasr A. (2015). Neonatal infections in Saudi Arabia: Association with cytokine gene polymorphisms. Central-European Journal of Immunology, 40(1), 68-77.

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