Specific gene expression taqman assays

Manufactured by Thermo Fisher Scientific

The Specific Gene Expression TaqMan assays are a set of pre-designed and pre-optimized real-time PCR assays developed by Thermo Fisher Scientific. These assays target specific genes and allow for the quantification of gene expression levels in various sample types.

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Lab products found in correlation

Omniscript rt kit, by Qiagen (2 mentions) Superscript 4, by Thermo Fisher Scientific (1 mentions) 7500 fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Superscript 3 rt kit, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Rneasy kit, by Qiagen (1 mentions) Dnase, by Qiagen (1 mentions) 7300 real time pcr system, by Thermo Fisher Scientific (1 mentions)

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TaqMan Gene Expression Assays (5455 citations) TaqMan assays (1810 citations) Gene Expression Assays (111 citations) TaqMan expression assays (54 citations) Specific TaqMan® Gene Expression Assays (27 citations) Taqman gene assays (19 citations) Gene-specific Taqman Assays (7 citations) Specific TaqMan assays (6 citations) Gene specific Taqman Gene Expression assays (5 citations) TaqMan Gene Expression Assays specific PCR primers sets (3 citations)

4 protocols using specific gene expression taqman assays

Quantification of mtDNA and mRNA Levels

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For mtDNA relative quantification, SYBR Green real-time qPCR was performed using primers specific to a mouse mtDNA region in the COI gene. Primers specific to RNaseP, a single copy gene taken as a nuclear gene reference. All primers are listed in Table 1. Approximately 25 ng of DNA was used per reaction.
For the quantification of mRNA levels, cDNA was retrotranscribed from total RNA extracted using the Omniscript RT kit (Qiagen). For mitochondrial transcripts CoI and Nd4, specific primers (Table 1) were used as described above with SYBR Green chemistry. Expression was calculated using the ΔΔCt analysis using Gapdh as reference.
Specific Gene Expression TaqMan assays (Invitrogen) were used for Polg and Polg2. Expression was calculated using the ΔΔCt analysis using B2m as reference.

Silva-Pinheiro P., Pardo-Hernández C., Reyes A., Tilokani L., Mishra A., Cerutti R., Li S., Rozsivalova D.H., Valenzuela S., Dogan S.A., Peter B., Fernández-Silva P., Trifunovic A., Prudent J., Minczuk M., Bindoff L., Macao B., Zeviani M., Falkenberg M, & Viscomi C. (2021). DNA polymerase gamma mutations that impair holoenzyme stability cause catalytic subunit depletion. Nucleic Acids Research, 49(9), 5230-5248.

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Quantifying mtDNA and Mitochondrial Transcripts

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For mtDNA relative quantification, SYBR Green real-time qPCR was performed using primers specific to a mouse mtDNA region in the COI gene. Primers specific to RNaseP, a single copy gene taken as a nuclear gene reference. All primers are listed in Table 1. Approximately 25 ng of DNA was used per reaction.
For the quantification of mRNA levels, cDNA was retrotranscribed from total RNA extracted using the Omniscript RT kit (Qiagen). For mitochondrial transcripts CoI and Nd4, specific primers (Table 1) were used as described above with SYBR Green chemistry. Expression was calculated using the ∆∆Ct analysis using Gapdh as reference.
Specific Gene Expression TaqMan assays (Invitrogen) were used for Polg and Polg2. Expression was calculated using the ∆∆Ct analysis using B2m as reference.

Silva-Pinheiro P., Viscomi C., Reyes A., Tilokani L., Mishra A., Cerutti R., Li S., Ho D.H., Valenzuela S., Dogan A., Peter B., Fernández‐Silva P., Trifunović A., Prudent J., Minczuk M., Bindoff L.A., Macao B., Zeviani M., Falkenberg M., & Viscomi C. (2020). In vivo and in vitro mechanistic characterization of a clinically relevant PolγA mutation.

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Transcriptional Profiling of Eosinophilic Esophagitis

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Tissue and globin reduced whole blood RNA (100ng) from adult EoE and GERD patients was reverse transcribed into cDNA using SuperScriptIV reverse transcriptase (Life Technologies). Gene expression was detected using Taqman gene expression assays specific for the STAT1, STAT2, ADAR, IFIT1, GBP2, and RPL36A genes (Applied Biosystems) on a 7500 Fast Real-Time PCR System (Applied Biosystems). Samples were run in triplicate in a single experiment. Mean Ct values were normalized to expression of the housekeeping gene (RPL36A), and then expressed relative to a technical positive control sample included on each plate.

Ruffner M.A., Hu A., Dilollo J., Benocek K., Shows D., Gluck M., Spergel J.M., Ziegler S.F., Hill D.A, & Cerosaletti K. (2021). Conserved interferon signature between adult and pediatric eosinophilic esophagitis. Journal of immunology (Baltimore, Md. : 1950), 206(6), 1361-1371.

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Real-Time PCR of Macrophage Markers

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Total RNA was isolated from purified F4/80⁺ cells using TRIzol reagent (Invitrogen, Carlsbad, CA) and then DNase (Qiagen, Germantown, MD) treated to remove possible traces of contaminating DNA according to the manufacturer’s instructions. Total RNA was subsequently recovered using the Qiagen RNeasy kit. cDNA was synthesized from 1 μg total RNA using the oligo(dT) primer and reagents supplied in the SuperScript III RT kit (Invitrogen) according to the manufacturer’s instructions. The cDNA was used as a template for real-time PCR analysis using the TaqMan gene expression assay (Applied Biosystems, Foster City, CA) according to the manufacturer’s instructions. All real-time PCR reactions were performed using the 7300 real-time PCR system (Applied Biosystems). For each real-time PCR reaction, a master mix was prepared on ice with TaqMan gene expression assays specific for iNOS, IFN-γ, TNF-α, CXCL9, CXCL10, IRF-1, SOCS-1, IL-17A, Ym1, FIZZ1, Arg1, IL-4, IL-13, and CD206 (Applied Biosystems). TaqMan rodent GAPDH (Applied Biosystems) was used as an internal control. The thermal cycling parameters contained an initial denaturing cycle of 95°C for 10 min followed by 40 cycles of 95°C for 15s and 60°C for 60s. Results of the real-time PCR data were derived using the comparative Ct method to detect relative gene expression as described previously (26 (link)).

Wager C.M., Hole C.R., Wozniak K.L., Olszewski M.A, & Wormley FL J.r. (2014). STAT1 Signaling is Essential for Protection against Cryptococcus neoformans Infection in Mice. Journal of immunology (Baltimore, Md. : 1950), 193(8), 4060-4071.

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