Taqman gene expression master mix 2

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

Taqman Gene Expression Master Mix (2×) is a pre-formulated, ready-to-use solution for real-time quantitative PCR (RT-qPCR) gene expression analysis. It contains all the necessary components, including DNA polymerase, dNTPs, buffer, and a passive reference dye, to perform RT-qPCR reactions.

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) Gene Expression Master Mix (97 citations) Taqman Gene Expression Mix (12 citations) Master Mix II (4 citations) TaqMan Gene Expression Master Mix II with UNG (2 citations)

9 protocols using taqman gene expression master mix 2

Comprehensive qPCR Workflow for Gene Expression Analysis

Cited 2 times

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

RNA was extracted using QIAshredder (Qiagen) and RNeasy Kit (Qiagen) with on-column RNase-free DNase treatment (Qiagen) according to manufacturer’s protocol. 2 µg of RNA was reverse transcribed using SuperScript IV (Thermo Fisher) and manufacturers protocol. qPCRs for AR, ERα, PGR, RUFY3, HPRT1, TFF1, PPL, CDKN1A, PCDH19, NOVA1, TET3, Gapdh, Tff1, Rara, Nrip1, Hsd11b2, Ppl, Pip, Cdkn1a, Pmepa1 and Pcdh19 were performed using TaqMan probes (Thermo Fisher, see Supplementary Table 6) with Taqman Gene Expression Master Mix (2×) (Thermo Fisher) and diluted cDNA. qPCR was performed using the following cycling conditions: hold at 50 °C for 2 min and 95 °C for 10 min, initial denaturation at 95 °C for 10 min followed by 40 cycles of 95 °C for 15 s and 60 °C for 1 min. qPCRs for GREB1, HSD11B2, PSA and HPRT1 were performed using previously published primers (see Supplementary Table 7) with Power SYBR Green Master Mix (Thermo Fisher) and diluted cDNA [17 (link), 50 (link)–52 (link)]. qPCR was performed using the following cycling conditions: hold at 95 °C for 10 min, 40 cycles of 95 °C for 15 s 60 °C for 1 min, melt curve: 95 °C for 15 s, 60 °C for 30 s and 95 °C for 15 s. Data was acquired using StepOne Real-Time PCR System and software V 2.0 (Applied Biosystems) and expression of the target gene was normalized against HPRT1 or Gapdh.

de Nys R., van Eyk C.L., Ritchie T., Møller R.S., Scheffer I.E., Marini C., Bhattacharjee R., Kumar R, & Gecz J. (2024). Multiomic analysis implicates nuclear hormone receptor signalling in clustering epilepsy. Translational Psychiatry, 14, 65.

+ Open protocol

+ Expand

qPCR Analysis of FZD3 Expression

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

RNA was extracted using QIAshredder and RNeasy Kit (Qiagen) and oncolumn RNase-free DNase treatment according to manufacturer's protocol. 2 µg RNA was reverse transcribed using SuperScript IV (Thermo Fisher). qPCRs for HPRT1 (Life Technologies, 4325801) and FZD3 (ThermoFisher, Hs00907280_m1) were performed using TaqMan probes with Taqman Gene Expression Master Mix (2×) (Thermo Fisher) and diluted cDNA using the following cycling conditions: hold at 50 °C for 2 min and at 95 °C for 10 min followed by 40 cycles of 95 °C for 15 s and 60 °C for 1 min. Data was acquired using StepOne Real-Time PCR System and software V 2.0 (Applied Biosystems) and expression of the target gene was normalised against HPRT1. Graph displays the mean and standard deviation (SD). Statistical analysis was performed using unpaired t-test with Welch's correction and 95% confidence interval.

de Nys R., Gardner A., van Eyk C., Mincheva-Tasheva S., Thomas P., Bhattacharjee R., Jolly L., Martinez-Garay I., Fox I.W.J., Kamath K.S., Kumar R, & Gecz J. (2024). Proteomic analysis of the developing mammalian brain links PCDH19 to the Wnt/β-catenin signalling pathway. Molecular psychiatry.

+ Open protocol

+ Expand

Quantifying myomiR-206 expression using qPCR

Cited 1 time

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

Reverse transcription reactions for myomiR‐206 and U6 small nuclear RNA (Rnu6) were performed with 10 ng of total RNA using Taqman MicroRNA Reverse Transcription Kit (ThermoFisher Scientific) according to the manufacturer's directions. qPCR was carried out with Taqman Gene expression Master Mix (2×) (ThermoFisher Scientific), TaqMan gene expression assay (miR‐206, #000510; Rnu6, #001973) using cDNA in a 10 µl reaction volume. qPCR was performed using the QuantStudio3 (Applied Biosystems) qPCR system as described by the manufacturer. qPCR efficiency was calculated by linear regression during the exponential phase using LinRegPCR software v11.126 (Ruijter et al., 2009 (link)). The comparison of miRNAs expression between groups was determined following normalization with Rnu6. Relative quantification of miRNA expression was assessed by the ΔΔCT method relative to the control.

Valentino T., Figueiredo V.C., Mobley C.B., McCarthy J.J, & Vechetti IJ J.r. (2021). Evidence of myomiR regulation of the pentose phosphate pathway during mechanical load‐induced hypertrophy. Physiological Reports, 9(23), e15137.

+ Open protocol

+ Expand

TaqMan-Based Real-Time PCR Method

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

We used TaqMan (Life Technologies) method on Applied Biosystems 7500 Fast Real-Time PCR System. mRNA was extracted using TRIzol (Life Technologies) according to the manufacturer's protocol. mRNA (1 μg) was used for first-strand complementary DNA (cDNA) synthesis with SuperScript III Kit (Life Technologies) according to the manufacturer's protocol. The first strand cDNA was further diluted 10 times with ribonuclease-free water and stored at −80°C. For 20 μl of realtime PCRs, the following reagents were added: 10 μl of TaqMan Gene Expression Master Mix (2×) (Life Technologies), 1 μl of TaqMan Gene Expression Assay (20×), 5 μl of nuclease-fee water, and 4 μl of cDNA. This reaction mix was added to the wells as duplicates for each sample. Real-time PCR products were run on a 1% agarose gel to confirm the size of the products. All the real-time PCR primers and probes were designed to span an exon junction to eliminate the contamination of genomic DNA. TaqMan primers and probes were used for CD59, β-actin, HMG-CoA synthase and reductase, ABCG1, and ABCA1. Results were expressed as C_t values normalized to the housekeeping gene β-actin.

Emin M., Wang G., Castagna F., Rodriguez-Lopez J., Wahab R., Wang J., Adams T., Wei Y, & Jelic S. (2016). Increased internalization of complement inhibitor CD59 may contribute to endothelial inflammation in obstructive sleep apnea. Science translational medicine, 8(320), 320ra1.

+ Open protocol

+ Expand

TaqMan-Based Real-Time PCR Method

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

+ Open protocol

+ Expand

Quantitative RT-PCR for MRAS and IGF1R

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

We synthesized cDNA from total RNA using a SuperScript VILO cDNA Synthesis Kit (Life Technologies). Quantitative real‐time PCR was performed on an ABI 7900HT System (Life Technologies). PCR for MRAS and IGF1R was carried out using TaqMan Gene Expression Master Mix (2×) (Life Technologies, TaqMan Gene Expression Assays (20×) (Assay ID; Hs00171926_m1 and Hs00609566_m1, respectively; Life Technologies), and 10 ng cDNA in a final volume of 50 μL. ACTB was used as the endogenous control. ACTB PCR was carried out using TaqMan Gene Expression Master Mix (2×) (Life Technologies), 400 nmol/L forward primer (5′‐CCTCGCCTTTGCCGATC‐3′), 400 nmol/L reverse primer (5′‐CGAGCGCGGCGATATCA‐3′), 100 nmol/L fluorescein (FAM) probe (5′‐CCGCCGCCAGCTCACCATG‐3′), and 10 ng cDNA in a final volume of 50 μL. The 2^−ΔΔCt method of relative quantification was used.

Yasumoto M., Sakamoto E., Ogasawara S., Isobe T., Kizaki J., Sumi A., Kusano H., Akiba J., Torimura T., Akagi Y., Itadani H., Kobayashi T., Hasako S., Kumazaki M., Mizuarai S., Oie S, & Yano H. (2016). Muscle RAS oncogene homolog (MRAS) recurrent mutation in Borrmann type IV gastric cancer. Cancer Medicine, 6(1), 235-244.

+ Open protocol

+ Expand

Quantifying hTERT Expression in hUC-MSCs

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

RNA was extracted from SCM-expanded and SFM-expanded hUC-MSCs in SFM using the E.Z.N.A Total RNA Kit (Omega Bio-Tek, Norcross, GA, USA). cDNA was synthesized by M-MLV Reverse Transcriptase (Invitrogen, Carlsbad, CA, USA). TaqMan-based real-time quantitative PCR assay was used to analyze the expression of hTERT (the human telomerase catalytic subunit gene) [23 (link)]. For each PCR run, a 20 μl reaction mix was prepared with 10 μl TaqMan Gene Expression Master Mix (2×; Applied Biosystems, Warrington, UK), 1 μl of 10 μM upper primer, 1 μl of 10 μM lower primer, 1 μl of 10 μM probe, 1 μl of cDNA and 6 μl ddH₂0. The reaction mixes were then placed in Applied Biosystems real-time PCR System 7300 with the following thermal cycling conditions: 50°C for 2 minutes, 95°C for 10 minutes, 55 cycles including 95°C for 15 seconds and 60°C for 1 minute. RPLP0 was used as a reference gene. cDNA prepared from HeLa cells was used as a positive control.

Wang Y., Wu H., Yang Z., Chi Y., Meng L., Mao A., Yan S., Hu S., Zhang J., Zhang Y., Yu W., Ma Y., Li T., Cheng Y., Wang Y., Wang S., Liu J., Han J., Li C., Liu L., Xu J., Han Z.B, & Han Z.C. (2014). Human mesenchymal stem cells possess different biological characteristics but do not change their therapeutic potential when cultured in serum free medium. Stem Cell Research & Therapy, 5(6), 132.

+ Open protocol

+ Expand

ILK Signaling Gene Expression Analysis

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

After one week of culture (beginning from cell seeding), the cells were harvested and RNA was extracted using TRI Reagent Solution (Ambion Inc.). cDNA synthesis was performed using High Capacity cDNA Reverse Transcription Kit (Applied Biosystems Inc.). Gene expression analysis was carried out in a TaqMan Human ILK Signaling Array (Applied Biosystems Inc.) using 5 μl TaqMan Gene Expression Master Mix (2×) (Applied Biosystems Inc.), with 5 μl cDNA in each well. PCR reactions were monitored in real time using the ABI 7900HT Fast Real-time PCR system (Applied Biosystems Inc., Foster City, CA). The thermal cycling conditions for real-time PCR were 50 °C for 2 mins, then 95 °C for 20 secs, and 40 cycles of denature (95 °C, 3 secs) and annealing/extension (60 °C, 30 secs). Relative quantitation of gene expression was determined using the ΔΔCt method.

Hsueh Y.J., Huang S.F., Lai J.Y., Ma S.C., Chen H.C., Wu S.E., Wang T.K., Sun C.C., Ma K.S., Chen J.K., Lai C.H, & Ma D.H. (2016). Preservation of epithelial progenitor cells from collagenase-digested oral mucosa during ex vivo cultivation. Scientific Reports, 6, 36266.

+ Open protocol

+ Expand

Quantification of miRNA Expression by qRT-PCR

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

cDNA, previously pre-amplified, was diluted 1:10 in Tris-EDTA (TE) buffer 1× and added to a final qRT-PCR reaction volume of 20 µL, which contained TaqMan MicroRNA assay primers (Applied Biosystems) for each miRNA (Table S1), TaqMan Gene Expression Master Mix (2×) (Applied Biosystems), and nuclease-free water. The reaction was performed using ABI 7500 fast real-time PCR systems (Applied Biosystems) at 95 °C for 10 min and 40 cycles at 95 °C for 15 s and 60 °C for 60 s.
After validating the Ct mean of the housekeeping genes, U6snRNA, miR-16, and miR-1228, as reference miRNAs, the relative levels of each specific miRNA was calculated using the equation 2^-∆Ct, where ∆Ct = mean Ct_miRNA-mean Ct_{(miR-U6,16&1228)}, and Ct = threshold cycle.

Pastor-Navarro B., García-Flores M., Fernández-Serra A., Blanch-Tormo S., Martínez de Juan F., Martínez-Lapiedra C., Maia de Alcantara F., Peñalver J.C., Cervera-Deval J., Rubio-Briones J., García-Rupérez J, & López-Guerrero J.A. (2020). A Tetra-Panel of Serum Circulating miRNAs for the Diagnosis of the Four Most Prevalent Tumor Types. International Journal of Molecular Sciences, 21(8), 2783.

+ 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!