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Taqman fast universal pcr master mix for taqman assays

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TaqMan® Fast Universal PCR Master Mix is a ready-to-use solution for performing TaqMan assays. It contains all the necessary components for real-time PCR amplification and detection, including a hot-start DNA polymerase, dNTPs, and optimized buffer system.

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Lab products found in correlation

Stepone real time pcr system, by Thermo Fisher Scientific (5 mentions) Trifast, by Avantor (4 mentions) Revertaid reverse transcriptase, by Thermo Fisher Scientific (2 mentions) Cfx96 real time system, by Bio-Rad (2 mentions) Stepone, by Bio-Rad (1 mentions) Revertaid rt reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Tripure, by Roche (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions)

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TaqMan(R) Fast Universal PCR Master Mix for TaqMan assays TaqMan universal PCR master mix assays TaqMan Fast Universal PCR Mix TaqMan Universal PCR master TaqMan PCR assays Taqman universal mix Fast Taqman mix TaqMan Master Mix Universal Master Mix PCR Master Mix

5 protocols using taqman fast universal pcr master mix for taqman assays

1

Quantitative RT-PCR Analysis of Mouse Tissues

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Total RNA was isolated from mouse islets, liver and WAT with a Trizol extraction system (TriFast, PEQLAB GmbH, Erlangen, Germany). cDNA synthesis and quantitative RT-PCR was performed as previously described45 (link). The Applied Biosystems StepOne Real-Time PCR system (Applied Biosystems, CA) with TaqMan® Fast Universal PCR Master Mix for TaqMan assays (Applied Biosystems, CA) was used for analysis. Cyclophilin (PPIA) and β-Actin (ACTB) were used as internal housekeeping controls and the quantitative analysis was performed with the ΔΔCT method. The following TaqMan® Gene Expression Assays were used: Ppia (Mm03024003_g1), Actb (Mm00607939_s1), Il1b (Mm00434228), Il6 (Mm00446190), Tnf (Mm00443258_m1), Ccl2 (Mm00441242_m1), Il4 (Mm00445259_m1), Il10 (Mm00439614_m1), Tgfb (Mm01178820_m1), Cd68 (Mm03047343_m1), Emr1 (F4/80) (Mm00802529_m1), Itgam (CD11b) (Mm00434455_m1), Itgax (CD11c) (Mm00498698_m1), Mrc1 (CD206) (Mm00485148_m1), Arg1 (Mm00475988_m1).
He W., Yuan T., Choezom D., Hunkler H., Annamalai K., Lupse B, & Maedler K. (2018). Ageing potentiates diet-induced glucose intolerance, β-cell failure and tissue inflammation through TLR4. Scientific Reports, 8, 2767.
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2

Quantification of PHLPP1 and PHLPP2 mRNA

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Total RNA was isolated from cultured human or mouse islets or INS-1E cells using TriFast (PEQLAB Biotechnologie, Germany). cDNA synthesis (RevertAid reverse transcriptase, Thermo Fisher Scientific (TFS), MA, USA) and quantitative RT-PCR was performed as previously described (Ardestani et al., 2014 (link)). The Applied Biosystems StepOne Real-Time PCR system (Applied Biosystems, CA, USA) with TaqMan® Fast Universal PCR Master Mix for TaqMan assays (Applied Biosystems) were used for analysis. TaqMan® Gene Expression Assays were used for PHLPP1 (#Hs01597875_m1), PHLPP2 (#Hs00982295_m1), PPIA (#Hs99999904_m1), and TUBA1A (#Hs00362387_m1) for human, Phlpp1 (#Mm01295850_m1), Phlpp2 (#Mm01244267_m1), Ppia (#Mm03024003_g1), and Tuba1a (#Mm00846967_g1) for mouse, and Phlpp1 (#Rn00572211_m1), Phlpp2 (#Rn01431647_m1), Ppia (#Rn00690933_m1) and Tuba1a (#Rn01532518_g1) for rat. qPCR was performed and analyzed by the Applied Biosystems StepOne Real-Time PCR system. The ΔΔCT method was used to analyze the relative changes in gene expression.
Lupse B., Annamalai K., Ibrahim H., Kaur S., Geravandi S., Sarma B., Pal A., Awal S., Joshi A., Rafizadeh S., Madduri M.K., Khazaei M., Liu H., Yuan T., He W., Gorrepati K.D., Azizi Z., Qi Q., Ye K., Oberholzer J., Maedler K, & Ardestani A. (2021). Inhibition of PHLPP1/2 phosphatases rescues pancreatic β-cells in diabetes. Cell reports, 36(5), 109490.
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3

Quantitative RT-PCR analysis of human islet genes

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Total RNA was isolated from cultured human islets using TriFast™ (peqGOLD; Peqlab) or TriPure (Roche) for FAC-sorted cells according to the manufacturer’s instructions. 500ng to 1μg of RNA were reverse transcribed using the RevertAid RT Reverse Transcription Kit (ThermoFisher) according to the manufacturer’s protocol, including removal of genomic DNA with DNase I prior to reverse transcription. Quantitative RT-PCR was performed as previously described [31 (link)]. StepOne Real-Time PCR system (Applied Biosystems, CA, USA) or CFX96 Real Time System (BioRad, CA, USA) with TaqMan® Fast Universal PCR Master Mix for TaqMan assays (Applied Biosystems) were used for analysis. TaqMan® Gene Expression Assays were used for human LDH-A (#Hs01378790_g1), PPiA (#Hs99999904_m1), ACTB (#Hs01060665_g1), GCG (#Hs01031536_m1), and INS (#Hs02741908_m1). qPCR was performed and analyzed by the Applied Biosystems StepOne or the BioRad CFX96 Real-Time Systems. The ΔΔCT or ΔCT methods were used to analyze the relative changes in gene expression.
Sanchez P.K., Khazaei M., Gatineau E., Geravandi S., Lupse B., Liu H., Dringen R., Wojtusciszyn A., Gilon P., Maedler K, & Ardestani A. (2021). LDHA is enriched in human islet alpha cells and upregulated in type 2 diabetes. Biochemical and biophysical research communications, 568, 158-166.
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4

Real-Time PCR Analysis of Mouse and Human Immune Markers

Cited 2 times
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For mouse skin, total RNA was isolated as stated above. For human PBMCs, total RNA was isolated from PBMC pellets with a Trizol extraction system (TriFast, PEQLAB GmbH, Erlangen, Germany), cDNA synthesis and quantitative RT-PCR was performed as previously described69 (link),70 (link). The Applied Biosystems StepOne Real-Time PCR system (Applied Biosystems, Forster City, CA) and TaqMan® Fast Universal PCR Master Mix for TaqMan assays (Applied Biosystems) were used for analysis. β-actin and cyclophilin were used as internal housekeeping controls and the quantitative analysis was performed with the ΔΔCT method. The following TaqMan® Gene Expression Assays (Applied Biosystems) were used: mouse Actb (Mm00607939_s1), mouse Tnf (Mm00443258_m1), human TNF (Hs99999043_m1), human PPIA (Hs99999904_m1).
All pre-clinical experimental analyses were performed in a blinded manner.
Mirastschijski U., Schwab I., Coger V., Zier U., Rianna C., He W., Maedler K., Kelm S., Radtke A., Belge G., Lindner P., Stahl F., Scharpenberg M., Lasota L, & Timm J. (2020). Lung Surfactant Accelerates Skin Wound Healing: A Translational Study with a Randomized Clinical Phase I Study. Scientific Reports, 10, 2581.
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5

Quantification of PHLPP1 and PHLPP2 mRNA

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was isolated from cultured human or mouse islets or INS-1E cells using TriFast (PEQLAB Biotechnologie, Germany). cDNA synthesis (RevertAid reverse transcriptase, Thermo Fisher Scientific (TFS), MA, USA) and quantitative RT-PCR was performed as previously described (Ardestani et al., 2014 (link)). The Applied Biosystems StepOne Real-Time PCR system (Applied Biosystems, CA, USA) with TaqMan® Fast Universal PCR Master Mix for TaqMan assays (Applied Biosystems) were used for analysis. TaqMan® Gene Expression Assays were used for PHLPP1 (#Hs01597875_m1), PHLPP2 (#Hs00982295_m1), PPIA (#Hs99999904_m1), and TUBA1A (#Hs00362387_m1) for human, Phlpp1 (#Mm01295850_m1), Phlpp2 (#Mm01244267_m1), Ppia (#Mm03024003_g1), and Tuba1a (#Mm00846967_g1) for mouse, and Phlpp1 (#Rn00572211_m1), Phlpp2 (#Rn01431647_m1), Ppia (#Rn00690933_m1) and Tuba1a (#Rn01532518_g1) for rat. qPCR was performed and analyzed by the Applied Biosystems StepOne Real-Time PCR system. The ΔΔCT method was used to analyze the relative changes in gene expression.
Lupse B., Annamalai K., Ibrahim H., Kaur S., Geravandi S., Sarma B., Pal A., Awal S., Joshi A., Rafizadeh S., Madduri M.K., Khazaei M., Liu H., Yuan T., He W., Gorrepati K.D., Azizi Z., Qi Q., Ye K., Oberholzer J., Maedler K, & Ardestani A. (2021). Inhibition of PHLPP1/2 phosphatases rescues pancreatic β-cells in diabetes. Cell reports, 36(5), 109490.
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