Taqman pcr master mix 2x

Manufactured by Thermo Fisher Scientific

Sourced in United States

TaqMan PCR Master Mix 2X is a ready-to-use solution for real-time PCR amplification. It contains all the necessary components, including DNA polymerase, dNTPs, and buffer, to perform quantitative PCR reactions.

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Lab products found in correlation

Quantstudio 12k sequence detection system, by Thermo Fisher Scientific (2 mentions) Rneasy mini kit, by Qiagen (1 mentions) Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Oligo dt, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

TaqMan Master Mix (448 citations) PCR Master Mix (310 citations) TaqMan 2X Universal PCR Master Mix (80 citations) TaqMan Fast Universal PCR Master Mix (2X) (63 citations) PCR Master Mix (2X) (48 citations) TaqMan™ Fast Universal PCR Master Mix (2X) no AMPERASE™ UNG (7 citations) Taqman 2X universal PCR master mix (no AmpErase UNG) (4 citations) Taqman One-Step RT-PCR Master Mix 2X (3 citations) TaqMan Universal PCR Master Mix II (2x), (3 citations) TaqMan Master Mix (2x) (2 citations)

3 protocols using taqman pcr master mix 2x

Genomic DNA Extraction and Genotyping

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Five ml blood samples were collected from each patient, by venipuncture in Vacutainer EDTA tubes, which were used for the subsequent extraction of DNA.
The genomic DNA was isolated from peripheral blood leukocytes with the QIAamp DNA Mini kit, and the genotype analysis was carried out by the allelic discrimination test using TaqMan probes. Fluorescence was quantified for each sample in Step One equipment for allelic discrimination (Applied Biosystems Foster City, CA, USA).
The amplification assays of the allelic variants were performed as follows: 30 ng of DNA sample, 220 μl of TaqMan PCR Master Mix 2X, 0.4 μl of 20X of each probe (Applied Biosystems) and 300 μl of water. The allelic variants were characterized by the fluorescent 5’ exonuclease method (TaqMan) and then independently analyzed.

Espindola L.M., López M.D., Flores A.D., Espinosa L.R., Granados J., Pacheco J.L., García M.P., Cervantes M.T., Méndez V.C., Doño S.H, & Ruíz Gómez D. (2019). Genetic polymorphism of CYP3A4 is associated with poor response to ifosfamide treatment in children with solid embryonic tumors. Archives of Medical Science : AMS, 17(6), 1766-1771.

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Gene Expression Analysis of WSB1 and IL21R

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To evaluate the expression levels of WSB1 and IL21R genes, RNA was isolated from PBL cultures using RNeasy Mini Kit (Qiagen, Hamburg, Germany) and 0.3 μg of total RNA from each sample was reverse transcribed into cDNA using 200 U of Superscript III Reverse Transcriptase (Life Technologies, Carlsbad, CA, USA) and 500 ng of Oligo (dT) (Life Technologies, Carlsbad, CA, USA), as described previously (36 (link)). Pre-synthesized Taqman® Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) were used to amplify WSB1 (Hs00373204_m1), IL21R (Hs00222310_m1) and β-actin (Hs01060665_g1). cDNA was detected using QuantStudio 12K Sequence Detection System (Applied Biosystems, Foster City, CA, USA). Each qRT-PCR assay was performed with 10 ng of cDNA in 10 μL of Taqman-PCR Master mix 2X (Applied Biosystems, Foster City, CA, USA) and 1 μL of primer/probe set and purified using deionized H₂O q.s. 20 μL. Gene expression was normalized to β-actin levels. Relative quantification was performed using the comparative threshold cycle (ΔΔCT) method (37 (link)–39 (link)).

Carneiro V.L., da Silva H.B., Queiroz G.D., Veiga R.V., Oliveira P.R., Carneiro N.V., Pires A.D., da Silva R.R., Sena F., Belitardo E., Nascimento R., Silva M., Marques C.R., Costa R.D., Alcantra-Neves N.M., Barreto M.L., Cooper P.J, & Figueiredo C.A. (2021). WSB1 and IL21R Genetic Variants Are Involved in Th2 Immune Responses to Ascaris lumbricoides. Frontiers in Immunology, 12, 622051.

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Quantitative RT-PCR Analysis of Gene Expression

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Presynthesized Taqman® Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) were used to amplify the following sequences (Applied Biosystems primers/probes set numbers are shown in parentheses): DAD1 (Hs00154671_m1), OXA1L (Hs00192329_m1) and β-actin (Hs99999903_m1). cDNA samples derived from the investigated genes were detected by a QuantStudio 12 K Sequence Detection System (Applied Biosystems, Foster City, CA, USA), according to the manufacturer's recommendations. Each qRT-PCR assay was performed with 10 ng of the cDNA sample in 10 μL of Taqman-PCR Mastermix 2X (Applied Biosystems, Foster City, CA, USA) and 1 μL of the respective primer/probe set and was purified using deionized H2O q.s. 20 μL. The gene expression levels were normalized to β-actin levels. Relative quantification was performed by the comparative threshold cycle (ΔΔCT) method, as previously described (Hellemans et al., 2007; (link)Orlando et al., 1998; (link)Vandesompele et al., 2002) (link).

Pires A.O., Queiroz G.A., de Jesus Silva M., da Silva R.R., da Silva H.B.F., Carneiro N.V.Q., Fonseca H.F., de Santana M.B.R., Nascimento R.S., Alcântara-Neves N.M., Costa G.N.O., Costa R.D.S., Barreto M.L, & Figueiredo C.A. (2018). Polymorphisms in the DAD1 and OXA1L genes are associated with asthma and atopy in a South American population. Molecular immunology, 101.

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