Stepone with taqman pcr master mix

Manufactured by Thermo Fisher Scientific

Sourced in United States

The StepOne with Taqman PCR master mix is a real-time PCR system designed for gene expression analysis and quantification. It is capable of performing quantitative PCR (qPCR) reactions using Taqman probes. The system provides precise temperature control and fluorescence detection for accurate and reliable results.

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Lab products found in correlation

High pure rna isolation kit, by Roche (3 mentions) Gapdh, by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) High pure rna tissue kit, by Roche (2 mentions) Tnf α, by Thermo Fisher Scientific (1 mentions) Il 1β, by Thermo Fisher Scientific (1 mentions) Spred2, by Thermo Fisher Scientific (1 mentions) Rneasy ffpe kit, by Qiagen (1 mentions) High pure rna isolation, by Roche (1 mentions)

Spelling variants of this product

TaqMan Master Mix (448 citations) StepOne (326 citations) PCR Master Mix (310 citations)

4 protocols using stepone with taqman pcr master mix

RNA Extraction and RT-qPCR Analysis

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Total RNA was isolated from the cultured cells and whole lungs using a High Pure RNA Isolation Kit or High Pure RNA Tissue Kit (Roche Applied Science, Penzberg, Germany), respectively. First-strand cDNA was constructed from 1 µg total RNA using oligo (dT)_12–18 primers, and the cDNAs were used as templates for PCR. RT-qPCR analysis was performed using StepOne with Taqman PCR master mix (Applied Biosystems, Foster City, CA, USA). Primers (Spred-2, Mm01223872_g1) were purchased from Applied Biosystems. Gene expression was normalized using GAPDH expression as an internal control, and relative fold change values were calculated based on unstimulated or WT control group that were assigned an arbitrary value of 1.

Xu Y., Ito T., Fushimi S., Takahashi S., Itakura J., Kimura R., Sato M., Mino M., Yoshimura A, & Matsukawa A. (2014). Spred-2 Deficiency Exacerbates Lipopolysaccharide-Induced Acute Lung Inflammation in Mice. PLoS ONE, 9(10), e108914.

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Quantifying Nanog Expression in HSC-2 and OE-33 Cells

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HSC-2 and OE-33 cells were seeded at 5 × 10⁴ cells/mL in 6 well plates 24 h before treatment with different concentrations of DFX for 48 h. Total RNA was isolated from HSC-2 and OE-33 cells using Trizol Reagent (Gibco BRL, Grand Island, NY, USA) and High Pure RNA Isolation kit (Roche Applied Science, Basel, Switzerland), respectively. First-strand cDNA was constructed from total RNA using the oligo (dT) primer. Real-time quantitative PCR analysis was performed using StepOne with Taqman PCR master mix (Applied Biosystems, Foster City, CA, USA). The primers used in this study were: GAPDH (Applied Biosystems) and Nanog (Integrated DNA Technologies, Coralville, USA). The quantification of the gene of interest was normalized to GAPDH and expressed as fold-increases relative to the negative control for each treatment at each time point as previously described.

Katsura Y., Ohara T., Noma K., Ninomiya T., Kashima H., Kato T., Sato H., Komoto S., Narusaka T., Tomono Y., Xing B., Chen Y., Tazawa H., Kagawa S., Shirakawa Y., Kasai T., Seno M., Matsukawa A, & Fujiwara T. (2019). A Novel Combination Cancer Therapy with Iron Chelator Targeting Cancer Stem Cells via Suppressing Stemness. Cancers, 11(2), 177.

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Real-Time qPCR Analysis of Inflammatory Markers

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Total RNA was isolated from whole livers and cultured cells using Trizol Reagent (Gibco BRL, Grand Island, NY, USA) and High Pure RNA Isolation kit (Roche Applied Science, Basel, Switzerland), respectively. First-strand cDNA was constructed from total RNA using the oligo (dT)_12–18 primer. Real-time quantitative PCR analysis was performed using StepOne with Taqman PCR master mix (Applied Biosystems, Foster City, CA, USA). The primers used in this study were: GAPDH(Mn99999915_-g1), TNFα (Mn00443258_–m1), IL-1β (Mn00434228_–m1), FasL (Mm00438864.m1), and Spred2 (Mn01223872_-g1) (Applied Biosystems). The quantification of the genes of interests was normalized to GAPDH and expressed as fold-increases relative to the negative control for each treatment at each time point as previously described^{39 (link)}.

Yang X., Fujisawa M., Yoshimura T., Ohara T., Sato M., Mino M., San T.H., Gao T., Kunkel S.L, & Matsukawa A. (2018). Spred2 Deficiency Exacerbates D-Galactosamine/Lipopolysaccharide -induced Acute Liver Injury in Mice via Increased Production of TNFα. Scientific Reports, 8, 188.

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RNA Extraction and qPCR Analysis

Cited 1 time

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Total RNA was isolated from cultured cells and whole lungs using High Pure RNA Isolation and High Pure RNA Tissue kits (Roche Applied Science, Penzberg, Germany), respectively. RNA from human autopsy samples was isolated using an RNeasy FFPE kit (Qiagen, Germantown, MD). Total RNA was extracted, and 1µg of total RNA was reverse-transcribed to cDNA according to the procedure previously described. Real-time quantitative PCR analysis was performed using StepOne with Taqman PCR master mix (Applied Biosystems, Foster City, CA). The thermal cycling conditions included 50°C for 2 min and 95°C for 10 min, followed by 40 cycles of amplification at 95°C for 15 s and 55°C for 1.5 min for denaturing and annealing, respectively. Quantification of the genes of interests were normalized to GAPDH and expressed as fold increases over the negative control for each treatment at each time point, as previously described (7 (link)).

Ito T., Itakura J., Takahashi S., Sato M., Mino M., Fushimi S., Yamada M., Morishima T., Kunkel S.L, & Matsukawa A. (2016). Sprouty-related Ena/VASP homology 1-domain-containing protein-2 (Spred-2) critically regulates influenza A virus (H1N1)-induced pneumonia. Critical care medicine, 44(7), e530-e543.

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