Taqman gene expression master mix reagent

Manufactured by Thermo Fisher Scientific

Sourced in United States

TaqMan Gene Expression Master Mix is a ready-to-use reagent for real-time PCR gene expression analysis. It contains all the necessary components, including DNA polymerase, dNTPs, and reaction buffer, for efficient and sensitive target gene amplification and detection.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

TaqMan Gene Expression Master Mix (2233 citations) TaqMan Master Mix (448 citations) TaqMan reagents (162 citations) Gene Expression Master Mix (97 citations) Taqman Gene Expression Mix (12 citations) TaqMan gene expression reagents (12 citations) Taqman master mix reagent (7 citations)

10 protocols using taqman gene expression master mix reagent

Quantitative Analysis of RAI1 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated from subject or control EBV-immortalized lymphoblastoid cell lines using the RNeasy Mini-Kit (Qiagen, Valencia, CA). First strand cDNA was synthesized from DNase-treated RNA (Applied Biosystems, Austin, TX) using a high capacity RNA-to-cDNA kit (Applied Biosystems). qPCR was performed utilizing two RAI1 Assays-On-Demand Taqman primer-probe assays (Applied Biosystems), Hs00430773_m1 (Assay 1; exons 2–3 boundary) and Hs01554690_m1 (Assay 2; exons 3–4 boundary), and β-actin gene (Hs99999903_m1) for control. PCR amplifications were performed using 100 ng of cDNA using TaqMan Gene Expression Master Mix reagent (Applied Biosystems) and were carried out on an ABI PRISM 7900 HT Sequence Detection System (Applied Biosystems). Results were analyzed with the comparative CT method as described [Livak, 1997 ; Livak and Schmittgen, 2001 (link)].

Yeetong P., Vilboux T., Ciccone C., Boulier K., Schnur R.E., Gahl W.A., Huizing M., Laje G, & Smith A.C. (2016). Delayed Diagnosis in a House of Correction: Smith–Magenis Syndrome Due to a De Novo Nonsense RAI1 Variant. American journal of medical genetics. Part A, 170(9), 2383-2388.

+ Open protocol

+ Expand

Quantitative Analysis of RAI1 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated from patients’ or control lymphoblastoid cells using the RNeasy Mini-Kit (Qiagen, Valencia, CA). RNA was subsequently treated with DNA-free DNase (Applied Biosystems). RNA concentration and purity were assessed on a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE). First strand cDNA was synthesized using a high capacity RNA-to-cDNA kit (Applied Biosystems). qPCR was performed utilizing a RAI1 Assays-On-Demand Taqman primer-probe assay (Applied Biosystems), Hs00430773_m1 (located at the RAI1 exon 2–3 boundary) and a control assay for the beta-actin housekeeping gene (Hs99999903_m1). PCR amplifications were performed on 100ng of cDNA using TaqMan Gene Expression Master Mix reagent (Applied Biosystems) and were carried out on an ABI PRISM 7900 HT Sequence Detection System (Applied Biosystems). Results were analyzed with the comparative C_T method as described [10 , 11 ]. All assays were performed in triplicate. Displayed values in Figure 3 represent the relative quantification (RQ) normalized to the average of all control assays in all three control cell lines (arbitrarily set to 1).

Adams D.R., Yuan H., Holyoak T., Arajs K.H., Hakimi P., Markello T.C., Wolfe L.A., Vilboux T., Burton B.K., Fajardo K.F., Grahame G., Holloman C., Sincan M., Smith A.C., Wells G.A., Huang Y., Vega H., Snyder J.P., Golas G.A., Tifft C.J., Boerkoel C.F., Hanson R.W., Traynelis S.F., Kerr D.S, & Gahl W.A. (2014). Three Rare Diseases in One Sib Pair: RAI1, PCK1, GRIN2B Mutations Associated with Smith-Magenis Syndrome, Cytosolic PEPCK Deficiency and NMDA Receptor Glutamate Insensitivity. Molecular genetics and metabolism, 113(3), 161-170.

+ Open protocol

+ Expand

Quantifying RRAD mRNA Expression by RT-qPCR

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total cellular RNA was extracted using the RNeasy MiniKit (Qiagen, Valencia, CA) and was treated with DNase I (Qiagen). One microgram of RNA was converted to cDNA using an Omniscript RT Kit (Qiagen). Real-time PCR was performed using a Priam 7900HT Sequence Detection System (PE Applied Biosystems). RRAD mRNA and 18S rRNA were detected using TaqMan Gene Expression Master Mix Reagent and TaqMan probe (Applied Biosystems). Data were normalized to 18S rRNA as an endogenous control and were calculated using the comparative Ct method (2-delta delta Ct).

Kim H.K., Lee I., Kim S.T., Lee J., Kim K.M., Park J.O, & Kang W.K. (2019). RRAD expression in gastric and colorectal cancer with peritoneal carcinomatosis. Scientific Reports, 9, 19439.

+ Open protocol

+ Expand

Quantitative Analysis of PIGT Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary fibroblasts were cultured from forearm skin biopsies. Total RNA was isolated from control and patients’ fibroblasts, cultured in Dulbecco’s Modified Eagle Medium (DMEM), High Glucose (Invitrogen, Carlsbad, CA) to confluence in a 10-cm dish, using the RNeasy Mini-Kit (Qiagen, Valencia, CA) including on-column treatment with DNA-free DNase (Qiagen). 2μg of RNA were reverse transcribed with the Omniscript RT Kit (Qiagen) using random nonamer primers. qPCR was performed utilizing the PIGT Taqman Assay-On-Demand, Hs00988032_g1, located at the PIGT NM_015937.5 exon 4–5 boundary (Applied Biosystems, Foster City, CA), and the human GAPD (GAPDH) Endogenous Control (VIC®/TAMRA™ probe, primer limited; 4310884E, Life Technologies, Grand Island, NY). PCR amplifications were performed on 100 ng of cDNA using TaqMan Gene Expression Master Mix reagent (Applied Biosystems) and were carried out on an ABI 7500 FAST Real-Time PCR System (Applied Biosystems). Results were analyzed with the comparative C_T method using the manufacturer’s software (v.2.0.1) accompanying the ABI 7500 FAST Real-Time PCR System (Applied Biosystems). All assays were performed in triplicate.

Lam C., Golas G.A., Davids M., Huizing M., Kane M.S., Krasnewich D.M., Malicdan M.C., Adams D.R., Markello T.C., Zein W.M., Gropman A.L., Lodish M.B., Stratakis C.A., Maric I., Rosenzweig S.D., Baker E.H., Ferreira C.R., Danylchuk N.R., Kahler S., Garnica A.D., Schaefer G.B., Boerkoel C.F., Gahl W.A, & Wolfe L.A. (2015). Expanding the Clinical and Molecular Characteristics of PIGT-CDG, a Disorder of Glycosylphosphatidylinositol Anchors. Molecular genetics and metabolism, 115(2-3), 128-140.

+ Open protocol

+ Expand

Quantitative Expression Analysis of Laminin Genes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated from fibroblasts with TRIZOL reagent (Invitrogen) and from whole blood using PAXgene Blood RNA Kit (Qiagen). RNA was treated with a DNase kit (DNA-free), according to the manufacturer’s protocol (Applied Biosystems). RNA concentration and purity were assessed on a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific). First strand cDNA was synthesised using a high-capacity RNA-to-cDNA kit (Applied Biosystems). Human multiple tissue cDNA panels (human Multiple Tissue Panels of cDNA I and II and Human Fetal MT Panel) were purchased from Clontech. Quantitative real-time PCR (qPCR) was performed using a Bio-Rad iQ SYBR Green Supermix and qPCR machine with standard qPCR parameters to analyse the expression of LAMA1, LAMB1, LAMB2 and LAMC1 compared with the control gene ACTB (primer sequences available on request). For analysis of tissue-specific expression, LAMA1 (Hs01074511_m1), Assays-On-Demand primer-probe assays (Applied Biosystems) and a control assay for POL2RA (Hs00172187_m1) were used. PCR amplifications were performed on 100 ng of cDNA using TaqMan Gene Expression Master Mix reagent (Applied Biosystems) and were carried out on an ABI PRISM 7900 HT Sequence Detection System (Applied Biosystems). Results were analysed with the comparative C_T method as described.^{21 22 (link)}

Vilboux T., Malicdan M.C., Chang Y.M., Guo J., Zerfas P.M., Stephen J., Cullinane A.R., Bryant J., Fischer R., Brooks B.P., Zein W.M., Wiggs E.A., Zalewski C.K., Poretti A., Bryan M.M., Vemulapalli M., Mullikin J.C., Kirby M., Anderson S.M., Huizing M., Toro C., Gahl W.A, & Gunay-Aygun M. (2016). Cystic cerebellar dysplasia and biallelic LAMA1 mutations: a lamininopathy associated with tics, obsessive compulsive traits and myopia due to cell adhesion and migration defects. Journal of medical genetics, 53(5), 318-329.

+ Open protocol

+ Expand

HSV DNA Detection by TaqMan PCR

Check if the same lab product or an alternative is used in the 5 most similar protocols

HSV DNA was detected by the TaqMan real-time PCR method using a previously described primer set and probe (15 (link)). The primers amplify a 92 bp fragment within a highly conserved region of the DNA polymerase gene from the HSV-1 and HSV-2. Amplification was performed using TaqMan Gene Expression Master Mix Reagents (Applied Biosystems, Foster City, CA, USA) in an Applied Biosystems Sequence Detector 7500 machine (Applied Biosystems, Foster City, CA, USA).

Behzadi M.A., Davarpanah M.A., Namayandeh M., Pourabbas B., Allahyari S, & Ziyaeyan M. (2018). Molecular diagnosis of genital tract infections among HIV-positive women in Iran. Iranian Journal of Microbiology, 10(4), 233-241.

+ Open protocol

+ Expand

Real-Time PCR Detection of NG and CT

Check if the same lab product or an alternative is used in the 5 most similar protocols

Commercially available TaqMan real-time PCR kits were used to detect NG and CT in the clinical specimens. All the procedures were performed according to the directions and recommendations in the manufacturer’s protocols (Neisseria gonorrhoea and Chlamydia trachomatis standard kits; Primer Design Ltd., Millbrook Technology Campus, Southampton, UK). The amplification was performed using the TaqMan Gene Expression Master Mix Reagents (Applied Biosystems, Foster City, CA, USA) in a 7500 Real-Time PCR System instrument (Applied Biosystems, USA).

+ Open protocol

+ Expand

Quantitative Real-Time PCR Analysis of Gene Expression in MIN6 Cells and Islets

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA from MIN6 cells and islets was recovered using an RNeasy kit (Qiagen), reverse transcribed, and subjected to qRT-PCR using SyBR Green based methodology^{11 (link)}. Data for input RNA were normalized to TATA box binding protein (Tbp) or Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) message. Primer sequences were previously described for Insulin, Pdx1, Gck, Slc2a2, Ddit3, Atf4, Tbp, Gpx1, Nrf2, and Ppargc1a^{8 (link), 51 (link)–54 (link)}. Other primer sequences used were as follows: Block of proliferation 1 (BOP1): forward: 5′-GGCCCAACATGAATATGAAG-3′, reverse: 5′-TTGTAGATACGTTTGCCATC-3′; EBNA1 Binding Protein 2 (EBNA1BP2): forward: 5′-ATAAGCTGGATTTCCTGGAG-3′, reverse: 5′-ATTAGGCCCTTTACTCATCTG-3′; NOP56: forward: 5′-CCAGAGGAGTGTGAGGAGGTA-3′, reverse: 5′-GAGACAGGTGGGTCTTCCATTCC-3′; Proliferation-Associated 2G4 PA2G4: forward: 5′-GAAGGAGGGTGAATTTGTTG-3′, reverse: 5′-ATCTTGAACCTCCATCTCAG-3′. For Cdkn1a and gapdh, Taqman gene expression master mix reagents and gene expression assay probes were used (Applied Biosystems). Islets used for both immunoblot and PCR were isolated after 16 weeks of diet.

Hatanaka M., Anderson-Baucum E., Lakhter A., Kono T., Maier B., Tersey S.A., Tanizawa Y., Evans-Molina C., Mirmira R.G, & Sims E.K. (2017). Chronic high fat feeding restricts islet mRNA translation initiation independently of ER stress via DNA damage and p53 activation. Scientific Reports, 7, 3758.

+ Open protocol

+ Expand

Stem Cell Sphere Characterization via RT-qPCR

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were resuspended in a stem cell-permissive medium (DMEM/F12 containing 20 ng/mL EGF, bFGF, and N2 supplement (1×)). Spheres were imaged after 7 days using a microscope.
RNA isolation and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) RNA was extracted from cultured cells using RNeasy Puri cation Kit (Qiagen). Reverse transcription was performed using 1 µg of total RNA with SuperScript® IV First-Strand Synthesis System per the manufacturer's protocol (ThermoFisher). Target gene expression was determined using TaqMan primer and probe sets (Applied Biosystems) and TaqMan Gene Expression Master Mix Reagents. PCR was performed on an Applied Biosystems 7900HT Real-Time PCR System. ISL1, KLK2, KLK3, TMRPSS2, and IGF1R primers and probes were designed as per Applied Biosystems (Assay IDs: Hs00158126_m1, Hs00428383_m1, Hs02576345_m1, Hs00237175_m1, Hs00609566_m1). The experiment was performed in triplicates for each sample. All data were normalized to the expression of GAPDH (Assay ID: Hs02786624_g1). A comparative threshold method was used to quantify target gene expression.

Choi J.D., Kim T.J., Jeong B.C., Jeon H.G., Jeon S.S., Kang M.Y., Yeom S.Y., & Seo S.I. (2021). ISL1 Romotes Enzalutamide Resistance in Castration-resistant Prostate Cancer (CRPC) through Epithelial to Mesenchymal Transition (EMT).

+ Open protocol

+ Expand

qPCR Analysis of Cell Cycle Genes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Quantitative real-time polymerase chain reaction (qPCR) was performed as described previously. 21 Briefly, total RNA was isolated from clones stably transfected with shRBM5 plasmids or control vectors using RNAiso reagent (TaKaRa, Otsu, Japan). qPCR was performed with TaqMan probes for p21, p27, and GAPDH with TaqMan Gene Expression Master Mix reagents (Applied Biosystems, Carlsbad, CA, USA) according to the manufacturer's protocol. Relative gene expression was calculated using the cycle threshold method, and the expression of GAPDH was used as an internal control.

Kobayashi T., Ishida J., Shimizu Y., Kawakami H., Suda G., Muranaka T., Komatsu Y., Asaka M, & Sakamoto N. (2017). Decreased RNA-binding motif 5 expression is associated with tumor progression in gastric cancer. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 39(3).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!