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Qpcr reagents

Manufactured by Thermo Fisher Scientific
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QPCR reagents are a set of chemical compounds used in the process of quantitative polymerase chain reaction (qPCR), a molecular biology technique for amplifying and quantifying DNA or RNA sequences. These reagents enable the detection and measurement of specific genetic targets within a sample.

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Lab products found in correlation

Rna extraction kit, by Qiagen (2 mentions) Histamine eia kit, by Cayman Chemical (1 mentions) Interleukin 3 (il 3), by Thermo Fisher Scientific (1 mentions) Ab37291, by Abcam (1 mentions) Ab62331, by Abcam (1 mentions) Ly364947, by Cayman Chemical (1 mentions) Ecl kit, by Thermo Fisher Scientific (1 mentions) Metformin, by Merck Group (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Superoxide dismutase sod assay kit, by Beyotime (1 mentions) Malondialdehyde mda assay kit tba method, by Nanjing Jiancheng (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Amphotericin b, by Thermo Fisher Scientific (1 mentions) Irdye 800cw bonetag optical probe, by LI COR (1 mentions) Cc 2583, by Lonza (1 mentions) Divinylbenzene, by Merck Group (1 mentions) 2 2 azodiisobutyramidinedihydrochloride aiba, by Merck Group (1 mentions)

Close spelling variants of this product

Platinum SYBR Green qPCR SuperMix-UDG reagents (43 citations) SYBR green qPCR reagent (9 citations) SYBR® GreenER™ qPCR reagent system (8 citations) SYBR Green qPCR SuperMix-UDG reagents (4 citations) RT-qPCR reagents (3 citations) SYBR Green Real-Time qPCR reagents (3 citations) TaqMan qPCR reagents (3 citations) Luminaris Color HiGreen reagents qPCR Master Mix (3 citations) Platinum® SYBR® Green qPCR SuperMix UDG with ROX reagent (3 citations) Maxima SYBR Green/ROX qPCR Master Mix reagents (2 citations)

11 protocols using qpcr reagents

1

Mast Cell Culture and Characterization

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All chemicals were obtained from Sigma–Aldrich (St Louis, MO) unless indicated otherwise. COX-2 inhibitor (SC-236) and TGF-β inhibitor (LY364947), and histamine EIA kit were obtained from Cayman Chemicals (Ann Arbor, MI). PGE2 EIA was obtained from R&D (Minneapolis, MN). qPCR reagents were obtained from Invitrogen (Carlsbad, CA). Primers for qPCR were obtained from IDT (Coralville, IA). Antibodies for immunoblot analysis were obtained from Abcam (Cambridge, MA; histidine decarboxylase (HDC): Rabbit polyclonal #ab37291; COX-2: Rabbit monoclonal #ab62331). Mast cells were obtained by culturing murine bone marrow in 10% FBS and IL-3 (10 ng/mL, Peprotech, Rocky Hill, NJ) containing IMDM media for 4–8 weeks. Cell cultures contained MC populations (FcεR+, c-kit+) greater than 90% as measured by flow cytometry, using antibodies from eBioscience (San Diego, CA) and BD Biosciences (San Jose, CA). Cells used for in vitro studies were incubated in 1 mL of 10% FBS containing IMDM media supplemented with 10 ng/mL of IL-3 in 12 well plates.
Ocana J.A., Romer E., Sahu R., Pawelzik S.C., FitzGerald G.A., Kaplan M.H, & Travers J.B. (2018). Platelet-activating factor-induced reduction in contact hypersensitivity responses is mediated by mast cells via cyclooxygenase-2-dependent mechanisms. Journal of immunology (Baltimore, Md. : 1950), 200(12), 4004-4011.
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2

Metformin and Thioflavin T Assays

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Metformin and Thioflavin T (ThT) reagent were purchased from Sigma-Aldrich (Saint Louis, MO, USA). A BCA protein assay kit and superoxide dismutase (SOD) assay kit were obtained from Beyotime Biotechnology. The malondialdehyde (MDA) Assay Kit (TBA method) was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). qPCR reagents and ECL kit were purchased from Invitrogen. All the antibodies were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA).
Lu X.Y., Huang S., Chen Q.B., Zhang D., Li W., Ao R., Leung F.C., Zhang Z., Huang J., Tang Y, & Zhang S.J. (2020). Metformin Ameliorates Aβ Pathology by Insulin-Degrading Enzyme in a Transgenic Mouse Model of Alzheimer's Disease. Oxidative Medicine and Cellular Longevity, 2020, 2315106.
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3

Characterization of CagA-Induced Inflammatory Response in CASMCs

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Human coronary artery smooth muscle cells (CASMCs) from 3 donors were commercially obtained (CC-2583; Lonza, Basel, Switzerland). Cell culture reagents, i.e., Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), Penicillin/Streptomycin, Amphotericin B, l-glutamine, HEPES, and trypsin were obtained from ThermoFisher Scientific, Waltham, MA, USA. Recombinant CagA protein containing amino acids 918 to 1147 and an N-terminal His tag was purchased from Abcam, Cambridge, UK. Reagents for the isolation of total RNA were purchased from Qiagen, Hilden, Germany. qPCR reagents were purchased from ThermoFisher Scientific, Waltham, MA, USA. ELISA kits for human IL-1β (DY201) and IL-6 (DY206-05) were purchased from R&D Systems, Minneapolis, MN, USA. IRDye 800CW BoneTag Optical Probe was purchased from LI-COR Biosciences, Lincoln, NE, USA.
Sundqvist M.O., Wärme J., Hofmann R., Pawelzik S.C, & Bäck M. (2023). Helicobacter Pylori Virulence Factor Cytotoxin-Associated Gene A (CagA) Induces Vascular Calcification in Coronary Artery Smooth Muscle Cells. International Journal of Molecular Sciences, 24(6), 5392.
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4

Synthesis of Functionalized Polymers

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2-Vinylpyridine (97%,
2VP; Sigma-Aldrich,
U.K.) and divinylbenzene (80 mol % 1,4-divinyl content, DVB; Sigma-Aldrich,
U.K.) were passed through a column of activated basic alumina to remove
inhibitors and impurities before use. 2,2′-Azodiisobutyramidine
dihydrochloride (AIBA; 97%) was purchased from Sigma-Aldrich (U.K.)
and used as received. Aliquat 336 surfactant (Thermo Fisher, U.K.)
was used as received. Poly(ethylene glycol) methyl ether methacrylate
(PEGMA, average Mn 2000 g mol–1, Sigma-Aldrich, U.K.) was supplied as a 50 wt % solution in H2O. PCR reagents were used as received. DNA extraction and
purification kits (Qiagen, U.K.) were used as per manufacturer’s
instructions. qPCR reagents (Thermo Fisher, U.K.) and viral extraction
reagents (Invitrogen, U.K.) were used as received.
Trinh E., Batt L.J., Yue Q., Du R., Jones S.T, & Fielding L.A. (2024). Bridging Flocculation of a Sterically Stabilized Cationic Latex as a Biosensor for the Detection of Microbial DNA after Amplification via PCR. Biomacromolecules, 25(3), 1629-1636.
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5

Silk Fibers and Curcumin Bioactivity

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Partially degummed silk fibers were purchased from Xiehe Silk Incorporation, Shengzhou, Zhejiang province, China. Curcumin (Cat#C7727, >80% pure), 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO). mRNA isolation kits were purchased from Qiagen Inc (Valencia, CA). Cell medium ingredients and qPCR reagents were purchased from Life Technologies (Grand Island, NY).
Li C., Luo T., Zheng Z., Murphy A.R., Wang X, & Kaplan D.L. (2014). Curcumin-functionalized silk materials for enhancing adipogenic differentiation of bone marrow-derived human mesenchymal stem cells. Acta biomaterialia, 11, 222-232.
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6

qPCR for miR-17 expression in glioma

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A qPCR for miR-17 expression in glioma cell lines and tissues was performed using TaqMan miRNA Reverse Transcription kit according to the manufacturer’s instructions. Reactions were set up in a 384-well plate using Taqman Universal PCR Master Mix (Life Technology) and run in an ABI Prism7900HT according to the manufacturer’s instructions. All qPCR experiments were performed in triplicate and the small nucleolar RNA U6 expression level was used as an endogenous control. All qPCR reagents were purchased from Life Technologies.
Micale L., Fusco C., Fontana A., Barbano R., Augello B., De Nittis P., Copetti M., Pellico M.T., Mandriani B., Cocciadiferro D., Parrella P., Fazio V.M., Dimitri L.M., D’Angelo V., Novielli C., Larizza L., Daga A, & Merla G. (2015). TRIM8 downregulation in glioma affects cell proliferation and it is associated with patients survival. BMC Cancer, 15, 470.
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7

RNA Extraction and qPCR Normalization

Cited 1 time
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RNA was extracted using an RNA extraction kit (Qiagen, Chatsworth, CA). qPCR reagents were from Applied Biosystems (Foster City, CA). Ribosomal protein 36B4 was used to normalize gene expression (Li et al., 2013 (link)).
Li Y., Lei D., Swindell W.R., Xia W., Weng S., Fu J., Worthen C.A., Okubo T., Johnston A., Gudjonsson J.E., Voorhees J.J, & Fisher G.J. (2015). Age-associated increase of skin fibroblast-derived prostaglandin E2 contributes to reduced collagen levels in elderly human skin. The Journal of investigative dermatology, 135(9), 2181-2188.
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8

Vascular Cell Culture Protocols

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Unless otherwise stated, all reagents were purchased from Sigma-Aldrich (Dublin, IRL). Both human aortic endothelial cells (HAECs) and human aortic smooth muscle cells (HASMCs), as well as their respective growth media were purchased from Promocell GmbH (Heidelberg, Germany). Recombinant human RANKL and TRAIL were purchased from R&D Systems (Minneapolis, MN, USA). Tumor necrosis factor-alpha (TNF-α) was purchased from Merck Millipore (Danvers, MA, USA). Primers were sourced from Sigma Aldrich and Eurofins Genomics (Ebersburg, Germany). ELISA DuoSet kits and alkaline phosphatase (ALP) activity assay kits were purchased from R&D Systems and BioAssay Systems (Hayward, CA, USA), respectively, whilst qPCR reagents were purchased from Applied Biosystems/ThermoFisher Scientific (Paisley, UK).
Harper E., Rochfort K.D., Forde H., Davenport C., Smith D, & Cummins P.M. (2017). TRAIL attenuates RANKL-mediated osteoblastic signalling in vascular cell mono-culture and co-culture models. PLoS ONE, 12(11), e0188192.
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9

Quantification of MXD3 gene expression

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Cells were collected by centrifugation (200 xg, 7 minutes, room temperature) and resuspended (without washing) in lysis buffer. RNA was isolated using the RNeasy mini kit (Qiagen). Following purification, up to 7 μg total RNA were digested with DNAse using Turbo DNA-free (Ambion) according to the manufacturer’s instructions to remove traces of genomic DNA. Up to 1 μg of DNAse-treated RNA was reverse-transcribed into cDNA using the High Capacity RNA to cDNA kit (Applied Biosystems). Quantitative PCR (qPCR) was performed in a 7900HT system using a validated MXD3 Taqman assay (Hs01557564_m1, Applied Biosystems) and TaqMan master mix. Triplicate 12 μl reactions were performed with 50 ng cDNA template per reaction in 384-well plates. Negative controls included no-reverse transcriptase reactions and water-only reactions. Relative expression levels were calculated by the comparative CT method, using 18S RNA as an endogenous control, also run in triplicate with the appropriate negative controls. Data were analyzed using SDS and DataAssist software. All qPCR reagents, instrument and software were from Applied Biosystems.
Barisone G.A., Satake N., Lewis C., Duong C., Chen C., Lam K.S., Nolta J, & Díaz E. (2014). Loss of MXD3 Induces Apoptosis of Reh Human Precursor B Acute Lymphoblastic Leukemia Cells. Blood cells, molecules & diseases, 54(4), 329-335.
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10

RNA Extraction and qPCR Normalization

Cited 1 time
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RNA was extracted using an RNA extraction kit (Qiagen, Chatsworth, CA). qPCR reagents were from Applied Biosystems (Foster City, CA). Ribosomal protein 36B4 was used to normalize gene expression (Li et al., 2013 (link)).
Li Y., Lei D., Swindell W.R., Xia W., Weng S., Fu J., Worthen C.A., Okubo T., Johnston A., Gudjonsson J.E., Voorhees J.J, & Fisher G.J. (2015). Age-associated increase of skin fibroblast-derived prostaglandin E2 contributes to reduced collagen levels in elderly human skin. The Journal of investigative dermatology, 135(9), 2181-2188.
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