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Taqman fast advanced pcr master mix

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TaqMan™ Fast Advanced PCR Master Mix is a ready-to-use solution for real-time PCR applications. It contains all the necessary components, including DNA polymerase, dNTPs, and buffer, to perform fast and efficient PCR reactions.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (6 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (5 mentions) High capacity cdna archive kit, by Thermo Fisher Scientific (4 mentions) 7900ht fast real time pcr system, by Thermo Fisher Scientific (4 mentions) 7500 fast real time pcr system, by Thermo Fisher Scientific (4 mentions) Purelink rna mini kit, by Thermo Fisher Scientific (3 mentions) Taqman microrna assay, by Thermo Fisher Scientific (2 mentions) Taqman primer, by Thermo Fisher Scientific (2 mentions) Quantstudio 3 real time pcr system, by Thermo Fisher Scientific (2 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Iscript cdna synthesis kit, by Bio-Rad (2 mentions) Verso cdna synthesis kit, by Thermo Fisher Scientific (2 mentions) Taqman advanced mirna cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) Sds software, by Thermo Fisher Scientific (1 mentions) Cdna synthesis kit, by Bio-Rad (1 mentions) Quadromacs magnet, by Miltenyi Biotec (1 mentions) High capacity cdna rt kit, by Thermo Fisher Scientific (1 mentions) Genelute mammalian total rna purification kit, by Merck Group (1 mentions) Picopure rna isolation kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

TaqMan® Fast Advanced Master Mix (7500 Real-Time PCR System, (2 citations) TaqMan Fast Advanced Master Mix on a StepOne Plus real-time PCR System (3 citations) TaqMan Fast Advanced Master Mix (1900 citations) PCR Fast Advanced Master Mix (4 citations) Taqman Fast Advanced Mix (9 citations) Fast Advanced Master Mix (75 citations) Fast Taqman mix (4 citations) TaqMan Master Mix (448 citations) PCR Master Mix (310 citations)

23 protocols using taqman fast advanced pcr master mix

1

Quantification of miRNAs using TaqMan assays

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Total RNA was reverse transcribed using TaqMan advanced miRNA cDNA synthesis kit (Life Technologies, Waltham, MA, USA). Quantification of miRNAs was completed using the TaqMan fast advanced PCR master mix in conjunction with TaqMan miRNA expression assays (Life Technologies, Waltham, MA, USA). For miRNA normalization, miR-191-5p was used as an endogenous control [23 (link), 24 (link)]. The ΔΔCt method was used for analysis of real-time data to obtain a miRNA fold-change relative to miR-191-5p expression. Mann-Whitney-U was used for statistical analysis.
Awamleh Z., Gloor G.B, & Han V.K. (2019). Placental microRNAs in pregnancies with early onset intrauterine growth restriction and preeclampsia: potential impact on gene expression and pathophysiology. BMC Medical Genomics, 12, 91.
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2

Reverse Transcription and qPCR Analysis

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Total RNA (100 ng) was reverse-transcribed to cDNA using a High Capacity cDNA Archive Kit (LIFE TECHNOLOGIES; Carlsbad, CA, USA). TaqMan PCR was carried out using the TaqMan Fast Advanced PCR master mix and TaqMan gene expression assays (all reagents from LIFE TECHNOLOGIES), by means of a 7900HT Fast Real-Time PCR System (APPLIED BIOSYSTEMS) as previously reported40 (link).
Salati S., Genovese E., Carretta C., Zini R., Bartalucci N., Prudente Z., Pennucci V., Ruberti S., Rossi C., Rontauroli S., Enzo E., Calabresi L., Balliu M., Mannarelli C., Bianchi E., Guglielmelli P., Tagliafico E., Vannucchi A.M, & Manfredini R. (2019). Calreticulin Ins5 and Del52 mutations impair unfolded protein and oxidative stress responses in K562 cells expressing CALR mutants. Scientific Reports, 9, 10558.
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3

Pituitary Gland Gene Expression Analysis

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We performed qRT-PCR following the manufacturer's instructions in the 7500 Fast Real-Time PCR system (Life Technologies, CA, USA) and used TaqMan Fast Advanced PCR Master Mix and assays based on hydrolysis probes (TaqMan Gene Expression Assays, Life Technologies, CA, USA). We selected the following assays: GH (1573905 A6), FSHB (Hs00174919_m1), LHB (Hs00751207_s1), TSHB (Hs02759015_s1), PRL (Hs00168730_m1), POMC (Hs01596743_m1), AVPR1B (Hs00949767_m1) and CRHR1 (Hs00366363_m1). Reference genes used were: PGK1 (Hs00943178_g1), TBP (Hs00427620_m1) and MRPL19 (Hs00608519_m1). As a positive control, we used total RNA from normal adult brain tissue (Bionova). A pool of RNA from nine normal pituitary samples obtained from autopsies served as a calibrator. All samples were analysed in duplicate. The relative differences in gene expression were expressed as fold change and were obtained with the 2-ΔΔCt method (SDS software, Applied Biosystems).
García-Martínez A., Sottile J., Fajardo C., Riesgo P., Cámara R., Simal J.A., Lamas C., Sandoval H., Aranda I, & Picó A. (2018). Is it time to consider the expression of specific-pituitary hormone genes when typifying pituitary tumours?. PLoS ONE, 13(7), e0198877.
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4

Quantitative gene and miRNA expression

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Total RNA (100 ng) was reverse-transcribed to cDNA using a High Capacity cDNA Archive Kit (Life technologies; Carlsbad, CA, USA). TaqMan PCR was carried out using the TaqMan Fast Advanced PCR master mix and TaqMan gene expression assays (all reagents from Life Technologies), by means of a 7900HT Fast Real-Time PCR System (Applied Biosystems). Assays were performed in triplicate. Gene expression profiling was achieved using the comparative cycle threshold (CT) method of relative quantitation using Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as housekeeping genes. To normalize data, ΔΔCT was calculated for each sample using the mean of its ΔCT values subtracted from the mean ΔCT value measured in the control sample, set as a calibrator; relative quantitation (RQ) value was expressed as 2−ΔΔCT.
Individual miRNA detection by RT-qPCR was performed using the TaqMan MicroRNA assays (Life technologies). Individual reverse transcription reactions (5 ng of total RNA each target) were performed using the Taqman microRNA Reverse Transcription Kit and the miRNA-specific looped-primers. TaqMan PCR was performed in triplicate by using the 7900HT Fast Real-Time PCR System (Applied Biosystems). miRNA expression RQ data were calculated as reported above, using U6 snRNA as housekeeping control.
Salati S., Salvestrini V., Carretta C., Genovese E., Rontauroli S., Zini R., Rossi C., Ruberti S., Bianchi E., Barbieri G., Curti A., Castagnetti F., Gugliotta G., Rosti G., Bergamaschi M., Tafuri A., Tagliafico E., Lemoli R, & Manfredini R. (2017). Deregulated expression of miR-29a-3p, miR-494-3p and miR-660-5p affects sensitivity to tyrosine kinase inhibitors in CML leukemic stem cells. Oncotarget, 8(30), 49451-49469.
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5

Comprehensive gene and miRNA expression analysis

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Total RNA (100 ng) was reverse-transcribed to cDNA using a High Capacity cDNA Archive Kit (Life Technologies). TaqMan PCR was carried out by using the TaqMan Fast Advanced PCR master mix and TaqMan gene expression assays (all reagents from Life Technologies), by means of a 7900HT Fast Real-Time PCR System (Applied Biosystems). Assays were performed in triplicate. Gene expression profiling was achieved using the comparative cycle threshold (CT) method of relative quantitation using hypoxanthine phosphoribosyltransferase 1 (HPRT1) as the housekeeping gene. To normalize data, ΔΔCT was calculated for each sample using the mean of its ΔCT values subtracted from the mean ΔCT value measured in the MOCK sample, set as a calibrator; relative quantitation (RQ) value was expressed as 2−ΔΔCT.
Individual miRNA detection by RT-qPCR was performed using the TaqMan MicroRNA assays (Life technologies). Individual reverse transcription reactions (5 ng of total RNA each target) were performed using the Taqman microRNA Reverse Transcription Kit and the miRNA-specific looped-primers. TaqMan PCR was performed in triplicate by using the 7900HT Fast Real-Time PCR System (Applied Biosystems). miRNA expression RQ data were calculated as reported above, by using SNORD49A as the housekeeping control.
Bianchi E., Bulgarelli J., Ruberti S., Rontauroli S., Sacchi G., Norfo R., Pennucci V., Zini R., Salati S., Prudente Z., Ferrari S, & Manfredini R. (2015). MYB controls erythroid versus megakaryocyte lineage fate decision through the miR-486-3p-mediated downregulation of MAF. Cell Death and Differentiation, 22(12), 1906-1921.
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6

Quantitative Real-Time PCR for Gene Expression

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Total RNA (100 ng) was reverse transcribed to cDNA using a High Capacity cDNA Archive Kit (Life technologies; Carlsbad, CA). TaqMan PCR was carried out using the TaqMan Fast Advanced PCR master mix and TaqMan gene expression assays (all reagents from Life Technologies) by means of a 7900HT Fast Real-Time PCR System (Applied Biosystems). Assays were performed in triplicate. Gene expression profiling (GEP) was achieved using the comparative cycle threshold (CT) method of relative quantitation using Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as housekeeping genes. To normalize data, DDCT was calculated for each sample using the mean of its DCT values subtracted from the mean DCT value measured in the control sample, set as a calibrator; relative quantitation (RQ) value was expressed as 2 -DDCT .
Salati S., Prudente Z., Genovese E., Pennucci V., Rontauroli S., Bartalucci N., Mannarelli C., Ruberti S., Zini R., Rossi C., Bianchi E., Guglielmelli P., Tagliafico E., Vannucchi A.M, & Manfredini R. (2018). Calreticulin Affects Hematopoietic Stem/Progenitor Cell Fate by Impacting Erythroid and Megakaryocytic Differentiation. Stem cells and development, 27(4).
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7

Quantification of gene expression in HEK293 cells

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Total RNA was isolated from HEK293 cells using TRIzol solution following the indicated treatments. Total RNA (1 μg) was used to prepare cDNA with a cDNA synthesis kit (Bio-Rad, Hercules, CA) as per the company’s instructions. RT-PCR reactions were run on a 7500 Fast Real-Time PCR system (Applied Biosystems, Foster City, CA) using TaqMan Fast Advanced PCR master mix and TaqMan primers (Thermo Fisher Scientific). The mRNA expression levels of various genes are presented as fold change after normalization to an endogenous control GAPDH. The list of primers used in this study is provided in Table S1 (online only).
Srivastava R.K., Muzaffar S., Khan J., Traylor A.M., Zmijewski J.W., Curtis L.M., George J.F., Ahmad A., Antony V.B., Agarwal A, & Athar M. (2020). Protective role of HO-1 against acute kidney injury caused by cutaneous exposure to arsenicals. Annals of the New York Academy of Sciences, 1480(1), 155-169.
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8

Isolation and qPCR Analysis of Immune Cells

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Tumour tissues were carefully isolated 24 h after the final treatment (Day 20) and dissociated as described above. Macrophages or DCs were isolated by using F4/80 or CD11c Microbeads, respectively, on a QuadroMACS magnet (Miltenyi) according to the manufacturer’s instructions. Total RNA was extracted from the obtained cell populations by using a PureLink RNA Mini Kit (Applied Biosystems), and cDNA was prepared with High-Capacity cDNA RT Kit (ThermoFisher) according to the manufacturer’s instructions. RT-qPCR was done with the 7500 FAST Real-time PCR System (Applied Biosystems), with TaqMan™ Fast Advanced PCR master Mix (ThermoFisher) and TaqMan FAM dye-labelled probes including GAPDH (Hs02786624_g1, Mm99999915_g1), IFNA1 (Mm03030145_gH), and IFNB1 (Mm00439552_s1) (ThermoFisher). GAPDH was used to normalise signal expression. Fold-change comparisons were done between control and treated samples by using the ΔΔCT method as described elsewhere49 .
Lu Y., Huntoon K., Lee D., Wang Y., Ha J., Qie Y., Li X., Schrank B.R., Dong S., Gallup T.D., Kang M., Zhao H., An Y., Yang Z., Li J., Kim B.Y, & Jiang W. (2022). Immunological conversion of solid tumours using a bispecific nanobioconjugate for cancer immunotherapy. Nature nanotechnology, 17(12), 1332-1341.
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9

RNA Extraction and qRT-PCR Analysis

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RNA extraction from sorted cells was performed with PicoPure RNA Isolation Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Whole liver samples were homogenized by TissueLyzer (Retsch) in lysis solution and RNA extraction was performed with GenElute Mammalian Total RNA Purification Kit (Sigma) according to the manufacturer’s instructions. Total RNA was reverse transcribed into cDNA using the QuantiTect Reverse transcription kit (Qiagen) according to the manufacturer’s instructions. qRT-PCR reaction was performed with TaqMan Fast Advanced PCR Master Mix (Thermo Fisher Scientific) or Power SYBR Green PCR Master Mix (Thermo Fisher Scientific) and read by LightCycler 480 (Roche). Gene expression levels were calculated based on the ΔΔCt relative quantification method, normalized to Actb expression.
Inverso D., Shi J., Lee K.H., Jakab M., Ben-Moshe S., Kulkarni S.R., Schneider M., Wang G., Komeili M., Vélez P.A., Riedel M., Spegg C., Ruppert T., Schaeffer-Reiss C., Helm D., Singh I., Boutros M., Chintharlapalli S., Heikenwalder M., Itzkovitz S, & Augustin H.G. (2021). A spatial vascular transcriptomic, proteomic, and phosphoproteomic atlas unveils an angiocrine Tie–Wnt signaling axis in the liver. Developmental Cell, 56(11), 1677-1693.e10.
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10

Quantitative RNA Expression Analysis

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RNA was isolated from snap frozen mouse tissues using Direct-zol RNA MiniPrep kit (cat#R2051 Zymo Research). cDNA was prepared using MultiScribe Reverse Transcriptase (cat#4311235 Thermo Fisher). cDNA (50 ng) was amplified for 40 cycles by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) using Dnm1 (DNM1, Mn00802468_m1), Dnm2 (DNM2, Mn00514582_m1), Dnm3 (DNM3, Mn00554098_m1), AAK1 (Mn01183675_m1) and GAPDH (Mn99999915_g1) primers, QuantStudio 3 Real-Time PCR System, and TaqMan Fast Advanced PCR Mastermix (cat#4444556 Thermo Fisher). All samples were analyzed at least in duplicate and normalized by GAPDH expression. The relative expression ratio per condition was calculated as described [27 (link)].
Tonello R., Anderson W.B., Davidson S., Escriou V., Yang L., Schmidt B.L., Imlach W.L, & Bunnett N.W. (2022). The contribution of endocytosis to sensitization of nociceptors and synaptic transmission in nociceptive circuits. Pain, 164(6), 1355-1374.
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