Taqman rt reagents kit

Manufactured by Thermo Fisher Scientific

The TaqMan RT Reagents Kit is a set of reagents designed for the reverse transcription and real-time PCR (RT-qPCR) of RNA samples. The kit includes all the necessary components for efficient and reliable conversion of RNA to cDNA and subsequent quantification of target gene expression.

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Lab products found in correlation

Quantitect sybr green pcr kit, by Takara Bio (2 mentions) Trizol reagent, by Takara Bio (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Nanodrop 2000, by Thermo Fisher Scientific (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) Viral rna extraction kit, by Qiagen (1 mentions) Sybr green, by Thermo Fisher Scientific (1 mentions) Pe cy7 labeled anti cd45 hi30, by BD (1 mentions) Rpa t4, by BD (1 mentions) Magvet universal isolation kit, by Thermo Fisher Scientific (1 mentions) Platinum hifi taq dna polymerase, by Thermo Fisher Scientific (1 mentions) Cfx connect rt pcr detection system, by Bio-Rad (1 mentions) Ssoadvanced universal probes supermix, by Bio-Rad (1 mentions) Ifn γ, by R&D Systems (1 mentions) Primepcr probe assay, by Bio-Rad (1 mentions)

5 protocols using taqman rt reagents kit

HIV Pathogenesis in Humanized Mice

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Successfully engrafted NRG mice were infected intra-peritoneally with 200,000 TCID₅₀ of HIV-1 (either NL4.3 HIVwt or NL4.3 HIVNefsmr5a). The infected mice were monitored for HIV replication in vivo by measuring human CD4⁺ cells in blood and plasma viremia every two weeks. Peripheral blood was analyzed for total human CD4⁺ cell percent by flow cytometry with PE-CY7-labeled anti-CD45 (HI30; BD Bioscience), APC-labeled anti-CD3 (HIT3A; BD Bioscience), and PE-labeled anti-CD4 (RPA-T4; BD Biosciences). Flow cytometry was performed on a Beckman Coulter FC-500.
To determine levels of HIV plasma viremia, viral RNA was extracted from the blood of infected mice using a Qiagen Viral RNA extraction kit (Qiagen, Valencia, CA). Reverse transcription was performed with oligo dT primers using a Taqman RT Reagents kit (Applied Biosystems, Carlsbad, CA). Quantitative-PCR was performed using SYBR Green (Applied Biosystems, Carlsbad, CA) and a probe and primer pair specific for the HIV pol gene: 5′-CTGGCTACTATTTCTTTTGCTA-3′ and 5′-TGGCATGGGTACCAGCACA-3′ and probe 5′-TTTATCTACTT-GTTCATTTCCTCCATTCCTT-3′ (IDT DNA Technologies, Coralville, IA). Quantitative-PCR was performed on an Applied Biosystems 7200 analyzer.

Konadu K.A., Anderson J.S., Huang M.B., Ali S.A., Powell M.D., Villinger F, & Bond V.C. (2015). Hallmarks of HIV-1 pathogenesis are modulated by Nef’s Secretion Modification Region. Journal of AIDS & clinical research, 6(7), 476.

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Quantitative RNA Expression Analysis

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Before RNA extraction, NP tissue was placed in a mortar containing liquid nitrogen and ground with a pestle. Total RNA of NP tissues and NP cells were extracted by Trizol reagent (Takara Biotechnology) according to the manufacturer's instructions. The RNA concentration was determined using spectrophotometry. cDNAs were reverse-transcribed with TaqMan RT Reagents Kit (Applied Biosystems). Quantitative PCR was carried out using a QuantiTect SYBR Green PCR Kit (Takara) in an Applied Biosystems 7500 System. GAPDH gene expression was used as an endogenous control. The 2−ΔΔCT method was used to analyze the relative fold changes of target genes. The sequences of primers were shown in Supplementary Table 1.

Wu T., Li X., Jia X., Zhu Z., Lu J., Feng H., Shen B., Guo K., Li Y., Wang Q., Gao Z., Yu B., Ba Z., Huang Y, & Wu D. (2021). Krüppel like factor 10 prevents intervertebral disc degeneration via TGF-β signaling pathway both in vitro and in vivo. Journal of Orthopaedic Translation, 29, 19-29.

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Quantitative Gene Expression Analysis

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Total RNA was isolated using Trizol reagent (Takara Biotechnology) according to the manufacturer’s instructions. cDNA was synthesized using TaqMan RT Reagents Kit (Applied Biosystems). PCR was performed using a QuantiTect SYBR Green PCR Kit (Takara) on an Applied Biosystems 7500 System. GAPDH was used as an internal reference. PCR reactions were performed in triplicate. Gene expression was quantified using the 2^−ΔΔct method. The primer sequences were summarized in Supplementary Table 1.

Jia X., Li L., Wang F., Xue Y., Wu T., Jia Q., Li Y., Wu C., Chen Y., Wu J., Su Y., Wang X., Zhuang T., Dong X., Ling J., Yuan J, & Li Q. (2022). DUB3/KLF4 combats tumor growth and chemoresistance in hepatocellular carcinoma. Cell Death Discovery, 8, 166.

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Quantitative RT-PCR for FMDV Genomic Detection

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Viral RNA was extracted using the MagVet™ Universal Isolation Kit (Thermo Fisher Scientific) on a KingFisher^TM Flex Robot (Life Technologies). Following RNA extraction, cDNA was generated by using the TaqMan RT reagents Kit (Thermo Fisher Scientific) and hexamer random primers. Platinum HiFi Taq DNA Polymerase (Invitrogen) and the following primer pair were used to amplify the region of viral genome containing the inserted Nluc gene and to provide template for sequencing: forward/5′-CCACTCGGGTGACTGAACTGC-3′ and reverse/5′-TGTCCTCGAGTGATGCCATG-3′.
One-step Callahan 3D quantitative real-time RT-PCR was performed according to the standard protocol of the World Reference Laboratory for FMDV (King et al., 2006 (link)).

Zhang F., Perez-Martin E., Juleff N., Charleston B, & Seago J. (2017). A replication-competent foot-and-mouth disease virus expressing a luciferase reporter. Journal of Virological Methods, 247, 38-44.

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APOBEC3B Expression in Bladder Cancer

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Four bladder cancer cell lines (RT-4, KU-19-19, HT-1376, and UM-UC-3) were cultured in standard media (McCoy’s, RPMI1640, EMEM, and EMEM, respectively) with 10% FBS and 1% penicillin/streptomycin (Thermo Fisher Scientific) with the addition of either IFNγ (10 ng/mL; R&D Systems) or the equivalent amount of sterile PBS for 48 h. RT-4, HT-1376, and UM-UC-3 were obtained from ATCC, KU-19-19 was obtained from DSMZ. All experiments were performed on passages 4–10. Cells were lysed with TRIzol (Thermo Fisher Scientific) and total RNA was isolated following standard Abcam RNA isolation protocol. Quality and quantity of isolated RNA was evaluated with NanoDrop 2000 (Thermo Fisher Scientific). cDNA was prepared using standard amounts of RNA per sample (600 ng; 250 ng/uL) with the TaqMan RT Reagents kit (Thermo Fisher Scientific). Samples were run in technical triplicate in a 20-μL SsoAdvanced Universal Probes Supermix (BioRad) in standard 96-well PCR plates. Expression level of APOBEC3B and endogenous control GAPDH was measured using BioRad PrimePCR Probe Assays (qHsaCIP0039581 and qHsaCEP0041396) using BioRad CFX Connect RT-PCR Detection System. Experiment was repeated in triplicate before analysis, and the ΔΔCt method was used to calculate differences in gene expression between IFNγ-stimulated cells and controls.

Glaser A.P., Fantini D., Wang Y., Yu Y., Rimar K.J., Podojil J.R., Miller S.D, & Meeks J.J. (2017). APOBEC-mediated mutagenesis in urothelial carcinoma is associated with improved survival, mutations in DNA damage response genes, and immune response. Oncotarget, 9(4), 4537-4548.

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