Universal taqman mastermix

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Universal TaqMan MasterMix is a pre-formulated, ready-to-use solution for real-time PCR amplification and detection of target DNA sequences. It contains all the necessary components, including DNA polymerase, dNTPs, and buffer, to perform qPCR reactions.

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Lab products found in correlation

Taqman gene expression assay, by Thermo Fisher Scientific (3 mentions) Ready to go you prime first strand beads, by GE Healthcare (3 mentions) Rneasy mini kit, by Qiagen (3 mentions) Taqman assay, by Thermo Fisher Scientific (3 mentions) Iscript reverse transcription supermix, by Bio-Rad (2 mentions) 7500 fast real time pcr system, by Thermo Fisher Scientific (2 mentions) Tissuelyser lt, by Qiagen (2 mentions) Experion rna stdsens analysis kit, by Bio-Rad (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) Reverse transcription system, by Promega (1 mentions) Specific taqman probes, by Thermo Fisher Scientific (1 mentions) 5 mm stainless steel bead, by Qiagen (1 mentions) High capacity reverse transcription kit with rnase inhibitor, by Thermo Fisher Scientific (1 mentions) Single cell to ct kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

TaqMan PCR Universal Mastermix (13 citations) TaqMan 2x Universal Mastermix (11 citations) FAM-labeled Taqman universal PCR mastermix (5 citations) TaqMan Fast Universal PCR Mastermix No AmpErase UNG (5 citations) Taqman Universal PCR Mastermix no UNG (4 citations) Taqman Universal Mastermix without UNG (3 citations) Taqman Fast Universal Mastermix No AmpErase® UNG (3 citations) Taqman Universal PCR Mastermix and Gene-expression Assays (3 citations) Fast Taqman Universal PCR Mastermix (2 citations) Universal TaqMan 2 × PCR mastermix (2 citations)

9 protocols using universal taqman mastermix

Mouse Transcriptome Profiling via Microarray

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RNAs were reverse-transcribed using iScript reverse transcription supermix (Bio-Rad Laboratories #170-8841, Hercules, CA). Transcriptome profiling was performed at the UNC Lineberger Comprehensive Cancer Center (LCCC) Genomic Core Facility using the Agilent Once Color 80k60k Sure Print G3 Mouse Gene Expression Array (G4858A-028005). TaqMan gene expression assays (Life Technologies) were performed using universal TaqMan master mix (Life Technologies #4304437). The method for RNA isolation and Real-time RT-PCR was described previously [19 (link)]. Primer pairs and probes for RT-qPCR assays are listed below:
F; 5′-Fluorescein (FAM)
Q; Quencher (TAMRA)

Willis M.S., Holley D.W., Wang Z., Chen X., Quintana M., Jensen B.C., Tannu M., Parker J., Jeyaraj D., Jain M.K., Wolfram J.A., Lee H.G, & Bultman S.J. (2017). BRG1 and BRM Function Antagonistically with c-MYC in Adult Cardiomyocytes to Regulate Conduction and Contractility. Journal of molecular and cellular cardiology, 105, 99-109.

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Quantitative Real-Time PCR Analysis

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Total RNA was isolated from whole colonic tissue samples (~120 mg) using TRI-zol (Invitrogen, Carlsbad, California) according to manufacturer’s instructions. The extracted RNA was quantified via spectrophotometry, and converted to cDNA using the Reverse Transcription System (Promega, Madison, WI). qPCR was then completed using a master mix containing 2× Universal TaqMan master mix (Life Technologies, Grand Island, New York), 0.9 μM (each) forward and reverse primers (see Table 5), and 0.250 μg sample cDNA. 18S was used as the housekeeping gene. The PCR was performed using a Prism 7000 sequence detection system with the following thermoprofile: 2 min at 50°C, 10 min at 95°C, and then 40 amplification cycles of 15 s at 95°C and 1 min at 60°C. The relative amount of mRNA was determined using the comparative cycle threshold (C_T) method as previously described [75 (link)]. Non-stressed HCC control samples were used as baseline controls and were set at a value of 1. All other samples are based on a fold change from these control samples. Primer sequences have been previously published and are listed in Table 5[76 (link)]. All groups were n = 3 – 5, using a single experiment without replicates.

Galley J.D., Nelson M.C., Yu Z., Dowd S.E., Walter J., Kumar P.S., Lyte M, & Bailey M.T. (2014). Exposure to a social stressor disrupts the community structure of the colonic mucosa-associated microbiota. BMC Microbiology, 14, 189.

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Quantitative Analysis of Gene Expression in Urothelial Cells

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Total RNA from cultured urothelial cells was extracted using the RNeasy Mini Kit (Qiagen, Antwerp, Belgium), following manufacturer’s protocol. RNA concentration was determined in a micro-volume spectrophotometer DropSense16 (Trinean NV, Gent, Belgium). cDNA synthesis was performed with 1μg of total RNA using the Ready-To-Go You-Prime First-Strand Beads (GE Healthcare, Diegem, Belgium). Quantitative PCR reactions (20 μl), containing 3 μl cDNA template (diluted 1:5), Universal TaqMan MasterMix (2x concentrated, Life Technologies), specific TaqMan probes (Table 1, 20× concentrated, Life Technologies) and H₂O, were performed with the 7500 Fast Real-Time PCR System (Life Technologies). Reactions were made using the following program: 50°C for 2 min and 95°C for 10 min, followed by 40 cycles of 95°C for 15 s and 60°C for 1 min. Non-template controls (NTCs) were used as negative controls in every experiment.

Alpizar Y.A., Uvin P., Naert R., Franken J., Pinto S., Sanchez A., Gevaert T., Everaerts W., Voets T., De Ridder D, & Talavera K. (2020). TRPV4 Mediates Acute Bladder Responses to Bacterial Lipopolysaccharides. Frontiers in Immunology, 11, 799.

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Cardiac Tissue RNA Extraction and qRT-PCR

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Cardiac tissues were homogenized using a TissueLyser LT (Qiagen N.V. #69980, Venlo, The Netherlands) according to the manufacturer’s protocols. Approximately 20–40 mg of apical ventricle was homogenized in 1 mL of Trizol (Life Technologies #15596-026, Carlsbad, CA) using a 5-mm stainless steel bead (Qiagen N.V. #69989). Chloroform (200 μL) was added, centrifuged at 12,000 g (15 min at 4°C), isopropanol (0.5 mL) was then added to the aqueous phase, centrifuged at 12,000 g (10 min at 4°C), and the resulting RNA pellet was washed with 1 mL of 75% ethanol, then centrifuged at 7500 g (5 min at 4°C). The resulting pellet was dried and resuspended in RNase-free water. RNA concentrations were then determined by UV spectroscopy (absorbance of 260–280 nm). RNAs (500 ng) were reverse-transcribed using iScript reverse transcription supermix (Bio-Rad Laboratories #170-8841, Hercules, CA). TaqMan gene expression assays were performed using universal TaqMan master mix (Life Technologies #4304437) using the fourteen primer pairs shown in Table 1.

Jensen B.C., Bultman S.J., Holley D., Tang W., de Ridder G., Pizzo S., Bowles D, & Willis M.S. (2017). Upregulation of autophagy genes and the unfolded protein response in human heart failure. International journal of clinical and experimental medicine, 10(1), 1051-1058.

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Cardiac Gene Expression Analysis

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After euthanasia by cervical dislocation, hearts were removed, washed in ice-cold PBS, immediately snap frozen in liquid nitrogen and kept at −80 °C until final processing. Total RNA was extracted using the RNeasy Mini Kit (Qiagen), following the manufacturer’s protocol. RNA concentration and quality were assessed using the Experion RNA StdSens Analysis Kit (Bio-Rad). For cDNA synthesis, Ready-To-Go You-Prime First-Strand Beads (GE Healthcare) were used based on the supplier’s protocol. Generated cDNAs were stored at −20 °C. qPCR reactions, composed of cDNA template, Universal TaqMan MasterMix (2× concentrated, Life Technologies), TaqMan assay (20× concentrated, Life Technologies) and H₂O, were performed with the 7500 Fast Real-Time PCR System (Life Technologies). mPGK1 and mTBP, selected using the geNorm application, were used as endogenous controls. TaqMan assays used: CACNA1H-Mm00445382_m1, TRPM4-Mm00613173_m1, ACTA1-Mm00808218_g1, ANP-Mm01255748_g1, RCAN1-Mm01213407_m1, MYH7-Mm00600555_m1.

Kecskés M., Jacobs G., Kerselaers S., Syam N., Menigoz A., Vangheluwe P., Freichel M., Flockerzi V., Voets T, & Vennekens R. (2015). The Ca2+-activated cation channel TRPM4 is a negative regulator of angiotensin II-induced cardiac hypertrophy. Basic Research in Cardiology, 110(4), 43.

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RT-qPCR Validation of RNA-seq Findings

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cDNA was synthesised from 500 ng of total RNA using the High‐Capacity Reverse Transcription kit with RNase Inhibitor (Life Technologies, N8080119). Semiquantitative PCR was performed using Universal TaqMan Master Mix or Universal TaqMan Advanced Master Mix (RNA‐seq validation only) (Life Technologies, 4364340 and 4444965) on an ABI 7500 cycler or a Quant Studio 7 Pro PCR cycler (RNA‐seq validation only) with TaqMan gene expression assays (Life Technologies, 4331182): Δ40p53 [as previously described [19 (link)]] and TP53 (Hs01034249_m1). Relative expression was determined using the 2^−ΔΔCt method [25 (link)] by normalising to the housekeeping gene β‐microglobulin (B2M) and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) as previously described [19 (link)]. Additionally, the expression of five differentially expressed genes identified by RNA‐seq (based on mean normalised counts and relevance) was confirmed by RT‐qPCR using the following probes: LRG1 (Hs00364835_m1), HYOU1 (Hs00197328_m1), UBE2QL1 (Hs00331876_m1); SERPINA5 (Hs04333915_m1) and PCDH7 (Hs05574398_g1).

Zhang X., Groen K., Morten B.C., Steffens Reinhardt L., Campbell H.G., Braithwaite A.W., Bourdon J, & Avery‐Kiejda K.A. (2021). Effect of p53 and its N‐terminally truncated isoform, Δ40p53, on breast cancer migration and invasion. Molecular Oncology, 16(2), 447-465.

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Trigeminal and DRG RNA Extraction

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Mice were killed in a CO₂ chamber and the trigeminal and DRG were carefully dissected41 (link). DRG from the different segments of the spinal cord were pooled. Total RNA from trigeminal ganglia and pooled DRG was extracted with RNeasy Mini Kit (Qiagen), following the manufacturer's protocol. Quality and concentration of the extracted RNA was assessed using the Experion RNA StdSens Analysis Kit (Bio-Rad), and samples with RQI values <6 were discarded. Further, cDNA was synthesized with the extracted RNA using Ready-To-Go You-Prime First-Strand Beads (GE Healthcare). Quantitative reverse transcription–PCR (RT–PCR) was then performed following the protocol described before using 2 μl cDNA template, Universal TaqMan MasterMix (2 × concentrated, Life Technologies), TaqMan assay (20 × concentrated, Life Technologies). The used primers are listed in Supplementary Table 1. Non-template controls were used as negative controls in every experiment. PGK1 and HPRT1 were used as endogenous controls. Data are expressed as relative expression of detected mRNA normalized to PGK1.

Ghosh D., Pinto S., Danglot L., Vandewauw I., Segal A., Van Ranst N., Benoit M., Janssens A., Vennekens R., Vanden Berghe P., Galli T., Vriens J, & Voets T. (2016). VAMP7 regulates constitutive membrane incorporation of the cold-activated channel TRPM8. Nature Communications, 7, 10489.

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Single-cell AMPK subunit expression analysis

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PCR was performed as previously described [26 (link)]. Cytoplasm was collected by a patch pipette under
visual guidance by applying light suction. RNA was reverse transcribed into cDNA
and preamplified using the Single Cell to CT kit (Life Technologies, CA, USA).
Relative expression levels of AMPK subunits were determined by real-time PCR,
using Universal TaqMan MasterMix (2x concentrated, Life Technologies) and TaqMan
assay (20x concentrated, Life Technologies) using the ΔCt method [27 (link), 28 (link)].

Barrett C.E., Jiang M., O’Flaherty B.G., Dias B.G., Rainnie D.G., Young L.J, & Menigoz A. (2022). Early life exposure to high fructose diet induces metabolic dysregulation associated with sex-specific cognitive impairment in adolescent rats. The Journal of nutritional biochemistry, 114, 109220.

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Cardiac Gene Expression Analysis

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Total RNA was isolated from cardiac tissue homogenized with a TissueLyser LT (Qiagen N.V. #69980, Venlo, The Netherlands) using
Trizol (Life Technologies #15596-026, Carlsbad, CA) according to the manufacturer’s instructions. TaqMan gene expression assays (Life Technologies) were performed using universal TaqMan master mix (Life Technologies #4304437) commercial probes, as described previously^{24 (link)}.

Bultman S.J., Holley D., de Ridder G., Pizzo S., Sidorova T.N., Murray K.T., Jensen B.C., Wang Z., Bevilacqua A., Chen X., Quintana M.T., Tannu M., Rosson G.B., Pandya K, & Willis M.S. (2016). BRG1 and BRM SWI/SNF ATPases Redundantly Maintain Cardiomyocyte Homeostasis by Regulating Cardiomyocyte Mitophagy and Mitochondrial Dynamics In Vivo. Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology, 25(3), 258-269.

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