Taqman primer probe mixes

Manufactured by Thermo Fisher Scientific

TaqMan primer/probe mixes are pre-designed and pre-optimized oligonucleotides used in real-time PCR assays to detect and quantify specific DNA or RNA sequences. These mixes contain forward and reverse primers, as well as a fluorescent probe, designed to target a specific gene or transcript of interest.

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Lab products found in correlation

Rneasy mini kit, by Qiagen (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Taqman gene expression master mix, by Thermo Fisher Scientific (2 mentions) Direct zol rna miniprep kit, by Zymo Research (2 mentions) Taqman gene expression cells to ct kit, by Thermo Fisher Scientific (1 mentions) Quantitect reverse transcription kit, by Qiagen (1 mentions) High capacity rna to dna kit, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Steponeplus, by Thermo Fisher Scientific (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Rneasy plus micro kit, by Qiagen (1 mentions) Facsaria, by BD (1 mentions) Qscript xlt one step rt qpcr toughmix reagent, by Quanta Biosciences (1 mentions) Qscript cdna synthesis kit, by Quantabio (1 mentions) Sybr green, by Bio-Rad (1 mentions) Rnaeasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

Taqman primer-probe mix Gene-specific TaqMan primer-probe mixes TaqMan gene-specific primer (6-carboxyfluorescein [FAM]/MGB)-probe mixes TaqMan gene specific primer-(FAM/MGB) probe mixes Primer-probe mixes 20X primer-probe mix TaqMan probe mix TaqMan primer probes Primer Mix TaqMan primers

9 protocols using taqman primer probe mixes

Transcriptional Profiling of PJI-Derived Leukocyte Subsets

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CD45⁺ leukocyte populations (CD33⁺HLA-DR⁻, CD33⁻HLA-DR⁻CD16⁻, and CD33-HLA-DR⁻CD16⁺) from human PJI tissues were purified by FACS, whereupon total RNA was immediately isolated using the TaqMan gene expression Cells-to-CT kit (Ambion, Austin, TX). qRT-PCR was performed using TaqMan primer/probe mixes (Applied Biosystems, Foster City, CA) for the following genes: indoleamine-pyrrole 2,3-dioxygenase-1 (IDO-1), Proteinase 3, inducible nitric oxide synthase (iNOS), arginase-1 (Arg-1), IL-10, CD206, TNF-α, cyclooxygenase-2 (COX2), and the housekeeping gene GAPDH. Results are reported as genes being expressed (+) or absent (−) in each of the CD45⁺ leukocyte populations.

Heim C.E., Vidlak D., Scherr T.D., Hartman C.W., Garvin K.L, & Kielian T. (2015). Interleukin-12 promotes myeloid-derived suppressor cell (MDSC) recruitment and bacterial persistence during S. aureus orthopedic implant infection. Journal of immunology (Baltimore, Md. : 1950), 194(8), 3861-3872.

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Quantitative PCR Analysis of Astrocyte and bEND.3 Cell mRNA Expression

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Astrocytes and bEND.3 cells were cultured as described above, and mRNA
was harvested after 7-10 days using the RNeasy Mini Kit (Qiagen 74104) according
to the manufacturer’s instructions. Equal amounts of isolated mRNA were
reverse transcribed into cDNA using QuantiTect Reverse Transcription kit (Qiagen
205311), and one tenth of this reaction was used per quantitative polymerase
chain reaction (qPCR). Quantitative PCR was performed using TaqMan primer-probe
mixes (Applied Biosystems, TaqMan gene expression assay numbers: Mm01275814_m1
for Slc1a2, Mm00600697_m1 for Slc1a3, and Mm99999915_g1 for Gapdh), and an
Applied biosystem quantflex 7 was used for detection. Gapdh mRNA was measured in
multiplex assays. There were no differences between samples in gapdh levels;
therefore, data generated from qPCR are presented as those normalized to Gapdh.
TaqMan gene expression assays have a PCR efficiency of 100% (±10%) as
validated by Applied Biosystems. To determine the relative expression levels,
the comparative C_T (ΔΔ C_t) method was used
(Livak and Schmittgen, 2001 (link); Schmittgen and Livak, 2008 (link)).

Martinez-Lozada Z, & Robinson M.B. (2020). Reciprocal communication between astrocytes and endothelial cells is required for astrocytic glutamate transporter 1 (GLT-1) expression. Neurochemistry international, 139, 104787.

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Quantitative RT-PCR Analysis of RNA

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RNA isolated using TRIzol (Life Technologies) was reverse transcribed to cDNA using the high-capacity cDNA reverse transcription kit (Applied Biosystems) for subsequent qRT-PCR analysis. We used Taqman primer/probe mixes predesigned by Integrated DNA Technologies and Taqman gene expression master mix (Applied Biosystems) to perform qRT-PCR. Data are presented as fold changes in RNA levels compared to control treatment, calculated following the 2^−ΔΔCt method.

Umar S., Palasiewicz K., Volin M.V., Romay B., Rahat R., Tetali C., Arami S., Guma M., Ascoli C., Sweiss N., Zomorrodi R.K., O’Neill L.A, & Shahrara S. (2021). Metabolic regulation of RA macrophages is distinct from RA fibroblasts and blockade of glycolysis alleviates inflammatory phenotype in both cell types. Cellular and molecular life sciences : CMLS, 78(23), 7693-7707.

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Quantitative RT-PCR Analysis of RNA

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Quantification of Gene Expression in Astrocytes

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Astrocytes and bEND.3 cells were cultured as described above, and mRNA was harvested after 14 days using the RNeasy Mini Kit (Qiagen, Hilden, Germany), according to the manufacturer’s instructions. Equal amounts of isolated mRNA were reverse transcribed into cDNA using a high capacity RNA-to-DNA kit (Applied Biosystems, Foster City, CA), and one tenth of this reaction was used per quantitative polymerase chain reaction (qPCR). Quantitative PCR was performed using Taqman primer-probe mixes (Applied Biosystems) and a Stratagene Mx3005 qPCR system was used for detection.

Lee M.L., Martinez-Lozada Z., Krizman E.N, & Robinson M.B. (2017). Brain endothelial cells induce astrocytic expression of the glutamate transporter GLT-1 by a Notch-dependent mechanism. Journal of neurochemistry, 143(5), 489-506.

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Gene Expression Analysis of Differentiated HIE

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Differentiated HIE monolayers were cultured in 3 conditions: in 96-well plates, in the mPC without flow, and in the mPC with flow (200 μL hr⁻¹) as described above. After 24 hours, the cells were washed 2x with PBS and lysed with Trizol reagent (Ambion). RNA was isolated with the Direct-Zol RNA MiniPrep Kit (Zymo Research) following manufacturer’s instructions. 2-3 wells were pooled for one biological sample. qScript XLT 1-step RT-qPCR ToughMix with ROX master mix (Quantabio) and Taqman primer-probe mixes (ThermoScientific) were used to perform RT-qPCR (StepOne Plus, Applied Biosystems). Probe information is in Supplemental Table 1. Gene expression levels were normalized to GAPDH and fold change was calculated with the 2^−ΔΔCt method.

Wilson R.L., Hewes S.A., Rajan A., Lin S.C., Bomidi C., Iida T., Estes M.K., Maresso A.W, & Jane-Grande-Allen K. (2021). A millifluidic perfusion cassette for studying the pathogenesis of enteric infections using ex-vivo organoids. Annals of biomedical engineering, 49(4), 1233-1244.

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Monocyte Gene Expression Profiling

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Monocytes (CD11b^highLy6G^-Ly6C⁺F4/80^-) from the soft tissue surrounding the infected knee were purified by sorting on a FACSAria (BD Biosciences; RRID:SCR_016695) using the antibody panel described above, whereupon RNA was immediately isolated using an RNeasy Plus Micro Kit (Cat #74034 Qiagen, Hilden, Germany). cDNA was synthesized using the iScript cDNA Synthesis Kit (Cat #1708891 Bio-Rad, Hercules, CA) and qPCR was performed using TaqMan primer/probe mixes (ThermoFisher) for the following genes: arginase-1 (Arg-1; Cat #Mm00475988_m1), IL-10 (Cat #Mm00439616_m1), triggering receptor expressed on myeloid cells 2 (Trem2; Cat #Mm00451744_m1), hypoxia-inducible factor-alpha (HIF-1α; Cat #Mm00468869_m1), inducible nitric oxide synthase (iNOS; Cat #Mm00440485_m1), TNF-α (Cat #Mm00443258_m1), Trem1 (Cat #Mm00451738_m1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Cat #Mm99999915_g1). Gene expression levels of monocytes from CTO treated mice were normalized to GAPDH and are presented as the fold-induction (2^-ΔΔCt) value relative to monocytes isolated from CT treated animals. In separate experiments, monocyte gene expression was evaluated by NanoString, using the nCounter PanCancer Mouse Immune Profiling kit (Cat #XT-CSO-MIP1-12; NanoString, Seattle, WA). Results are expressed as the fold-change in monocytes recovered from CTO vs. CT treated mice.

Yamada K.J., Heim C.E., Xi X., Attri K.S., Wang D., Zhang W., Singh P.K., Bronich T.K, & Kielian T. (2020). Monocyte metabolic reprogramming promotes pro-inflammatory activity and Staphylococcus aureus biofilm clearance. PLoS Pathogens, 16(3), e1008354.

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Quantifying Gene Expression Changes

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Total RNA was extracted using Direct-zol^™ RNA MiniPrep Kit (Zymo Research) according to the manufacturer’s protocol. PCR reactions were performed using the qScript XLT One-Step RT-qPCR ToughMix reagent with 6-carboxy-X-rhodamine (ROX) (Quanta Biosciences) in the StepOnePlus real-time PCR system. Fold changes in mRNA expression were determined using the delta-delta-Ct method relative control samples after normalization to the housekeeping gene glyceraldehyde3-phosphate dehydrogenase (GAPDH). The following TaqMan primer-probe mixes (Thermo Fisher Scientific) were used: GAPDH, IFNλ1, IFI44L, IP-10, LCT, MTTP, APOB.

Adeniyi-Ipadeola G.O., Hankins J.D., Kambal A., Zeng X.L., Patil K., Poplaski V., Bomidi C., Nguyen-Phuc H., Grimm S.L., Coarfa C., Crawford S.E., Blutt S.E., Speer A.L., Estes M.K, & Ramani S. (2023). Infant and Adult Human Intestinal Enteroids are Morphologically and Functionally Distinct. bioRxiv.

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Quantifying Gene Expression in Murine Livers

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Total RNA was extracted from murine livers using the RNA Easy mini kit (Qiagen, Cat# 74104) and complementary DNA was synthesized from 1 µg of mRNA with the qScript cDNA Synthesis kit (Quantabio, Cat# 95048), in accordance with the manufacturer’s instructions. qRT-PCR was performed on 2 µL of cDNA using either SYBR-Green (Biorad, Cat# 1725270) or Taqman primer/probe mixes (Thermo Fisher Scientific). A table of primers can be found in the supplementary materials. The reactions were performed following the manufacturer’s instructions. GAPDH was employed as a housekeeping gene and fold changes in gene expression were calculated using the standard ∆∆Ct method.

Levine J.A., Oleaga C., Eren M., Amaral A.P., Shang M., Lux E., Khan S.S., Shah S.J., Omura Y., Pamir N., Hay J., Barish G., Miyata T., Tavori H., Fazio S, & Vaughan D.E. (2021). Role of PAI-1 in hepatic steatosis and dyslipidemia. Scientific Reports, 11, 430.

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