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Rt2 sybr green pcr master mix

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The RT2 SYBR Green PCR Master Mix is a ready-to-use solution for real-time PCR applications. It contains all the necessary components, including SYBR Green I dye, for the amplification and detection of target DNA sequences.

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Lab products found in correlation

Rt2 first strand kit, by Qiagen (5 mentions) Rt2 profiler pcr array, by Qiagen (4 mentions) Applied biosystems 7300 real time pcr system, by Thermo Fisher Scientific (1 mentions) Quantitect primer assay, by Qiagen (1 mentions) Mysticq microrna qpcr assay primer, by Merck Group (1 mentions) Mysticq microrna cdna synthesis mix, by Merck Group (1 mentions) Abi 7500 real time pcr system, by Thermo Fisher Scientific (1 mentions) Cfx96, by Bio-Rad (1 mentions) Cfx384 pcr machine, by Bio-Rad (1 mentions) Pcr array kit, by Qiagen (1 mentions) Lightcycler 480, by Roche (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Col6a3, by MyBioSource (1 mentions) Rt2 profiler pcr array plate, by Qiagen (1 mentions) Rt2 first strand cdna synthesis kit, by Qiagen (1 mentions) 7900 ht real time cycler, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

SYBR Green (372 citations) PCR master mix (50 citations) SYBR Green Mix (45 citations) RT2 SYBR Green (23 citations) SYBR master mix (17 citations)

12 protocols using rt2 sybr green pcr master mix

1

Validation of Methylated Genes and miRNA Expression

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For the validation of methylated genes expression, we used primers for Socs1, Ep300, p16, Mgmt and Dapk (QuantiTect Primer Assay, Qiagen); for the evaluation of miR-466e-5p gene targets expression, we used primers for Zfp704, Cplx2 and Cnot7[41] (link), [42] (link); for the validation of miRNA microarrays, we used primers for miR-466e-5p, miR-21-3p and miR-185-3p (MystiCq microRNA qPCR Assay Primer, Sigma-Aldrich). Mouse liver RNA was reverse-transcribed using the RT2 first strand kit (SABiosciences, Frederick, MD, USA) for gene expression experiments and MystiCq microRNA cDNA Synthesis Mix (Sigma-Aldrich) for miRNA expression experiments. Then quantitative real-time PCR reactions were performed in triplicate with an Applied Biosystems 7300 Real-Time PCR system (Life Technologies), using the RT2 SYBR Green PCR Master Mix (SABiosciences) for gene expression experiments and the MystiCq microRNA SYBR Green qPCR ReadyMix for miRNA expression experiments, according to the manufacturers’ instructions.
Vares G., Wang B., Ishii-Ohba H., Nenoi M, & Nakajima T. (2014). Diet-Induced Obesity Modulates Epigenetic Responses to Ionizing Radiation in Mice. PLoS ONE, 9(8), e106277.
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2

Inflammatory Cytokine Profiling in Wound Biopsies

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RNA was extracted from wound biopsies collected before and after MDT utilizing the PerfectPure RNA Isolation Kit for Fibrous Tissue (5 Prime Inc., Gaithersburg, MD). The purity of the extracted RNA was evaluated with an RT2 Profiler PCR quality control kit (SABiosciences, Frederick, MD). The mRNA levels of 84 distinct inflammatory cytokines were assessed using a real‐time PCR array (Array #PAMM‐011; SABiosciences) focusing on the inflammatory pathway, following the manufacturer's guidelines. The RNA samples were converted to complementary DNA (cDNA) using the RT2 First Strand Kit (SABiosciences), and the cDNA samples were combined with RT2 SYBR Green PCR Master Mix (SABiosciences). The presence of inflammatory cytokines in the samples was determined through RT2 Profiler PCR arrays (SABiosciences), including cytokine genes IFNG, IL‐10, TGF‐β1 and TNF. The specificity of amplification for each sample was confirmed using dissociation curve analysis, and agarose gel electrophoresis validated the amplicon size. The relative expression levels of each gene were calculated using the Delta–Delta CT method, with GAPDH serving as the reference gene.
Sun X., Chen J., Li G., Wang L., Wang T, & Wang A. (2023). Maggot debridement therapy stimulates wound healing by altering macrophage activation. International Wound Journal, 21(3), e14477.
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3

Quantitative Reverse Transcription PCR Assay

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We designed primers for these 90 CTA and 5 housekeeping genes (data not shown) using Primer 3 software and selected those that contained at least 1 exon-exon junction to reduce the genomic DNA contamination. One pair of primers for each gene was added to each well of a 96-well plate to form the assay, comprising 90-CTA genes and 5 housekeeping genes, and one well as a blank control without any primer. qRT-PCR analysis of the gene expression was then performed using RT2-SYBR® Green PCR Master Mix (Qiagen, Valencia, CA, USA) with 40 cycles of 15 s at 95°C and 1 min at 58°C on an ABI 7500 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). Fluorescence data were collected at 58°C after each cycle. After the final cycle, melting curve analysis of all samples was conducted within the range of 58–95°C. The specificity of the PCR products was verified by the targeted product size using gel electrophoresis and melting curve analysis. The threshold cycle and 2−ΔΔCt method were used for calculating the relative amount of the target RNA using the average of the five house-keeping genes as internal control, according to user’s manual. The experiments were repeated in triplicate.
Wen J., Li H., Tao W., Savoldo B., Foglesong J.A., King L.C., Zu Y, & Chang C.C. (2014). High throughput quantitative reverse transcription PCR assays revealing over-expression of cancer testis antigen genes in multiple myeloma stem cell-like side population cells. British journal of haematology, 166(5), 711-719.
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4

Oxidative Stress Gene Expression Analysis in Jurkat Cells

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The Human Oxidative Stress Plus RT2 Profiler PCR Array (PAHS-065Z) was used to analyze mRNA levels of 84 key genes related to oxidative stress (antioxidants, ROS metabolism and pathway activity signature genes) in a 96-well format, according to the manufacturer’s instructions (Qiagen). WT or PARP1 KO Jurkat cells were treated with or without APO866 at 10 nM for 40 hours. Reactions were run on a Real-Time PCR system, Bio-Rad CFX96, using RT2 SYBR Green PCR master mix (Qiagen, Switerland). The thermocycler parameters were 95°C for 10 min, followed by 40 cycles of 95°C for 15 s and 60°C for 1 min. Relative changes in gene expression analysis were performed according to QIAGEN Web software using the ΔΔCT method with normalization of the raw data to 5 housekeeping genes. The expression data are presented as real change multiples. Genes with altered expression profile compared to control with fold change value ≥ ±1.5 and P < 0.05 were presented.
Cloux A.J., Aubry D., Heulot M., Widmann C., ElMokh O., Piacente F., Cea M., Nencioni A., Bellotti A., Bouzourène K., Pellegrin M., Mazzolai L., Duchosal M.A, & Nahimana A. (2019). Reactive oxygen/nitrogen species contribute substantially to the antileukemia effect of APO866, a NAD lowering agent. Oncotarget, 10(62), 6723-6738.
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5

Quantifying Calcium Channel Gene Expression

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In a 96-well PCR array plate, the cDNA sample and the specific primer pairs were added to the RT2 SYBR Green PCR Master Mix (QIAGEN Inc., California, USA). We initially screened Ca2+ channel gene expression using PCR array PAHS-036 (Qiagen Inc., California, USA) to assess the presence of 84 ion channel genes and receptors in our samples. Subsequently, to quantify the expression levels of Ca2+ channel genes, we used predesigned PrimeTime® qPCR specific primers (Integrated DNA Technologies, Iowa, US) for CACNA1B (primer 1: 5’-GGTCTCTGGTGGTTTTGTTCT -3’; primer 2: 5’-AGGAGATTTCCGTTGTGTGG -3’), CACNA1C (primer 1: 5’- GTCCGCTTCCAGATCTTCTTG-3’; primer 2: 5’-TGTTCAATGCCACCCTGTT -3’), CACNA1D (primer 1: 5’-GGAAGTCTGGTGCCTCTTG- 3’; primer 2: 5’-GCTCAATAAATGTTCGTGGATGA-3’) and the transcription factor genes, MYC (primer 1: 5’-CGTAGTCGAGGTCATAGTTCC-3’; primer 2: 5’-GCTGCTTAGACGCTGGATT-3’) and CREB1 (primer 1: 5’-ATAAAACTCCAGCGAGATCCG-3’; primer 2: 5’-GTTACAGCTGCATCTCCACT-3’), and the housekeeping gene HPRT1(primer 1: 5’-AGGTATGCAAAATAAATCAAGGTCAT -3’; primer 2: 5’-TCCTCCTGAGCAGTCAGC -3’) and reran the real-time PCR for each experiment in triplicate or quadruplicate.
Andres M.A., Cooke I.M., Bellinger F.P., Berry M.J., Zaporteza M., Rueli R.H., Barayuga S.M, & Chang L. (2015). Methamphetamine acutely inhibits voltage-gated calcium channels but chronically upregulates L-type channels. Journal of neurochemistry, 134(1), 56-65.
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6

Quantitative Real-Time RT-PCR for Antibacterial Response

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Quantitative real-time RT-PCR was performed as previously described 10 (link). For PCR arrays, RNA was converted to cDNA with RT2 First Strand kit (#330401, Qiagen). Mouse antibacterial response PCR arrays (PAMM-148Z) were performed with the RT2 SYBR Green PCR master mix (#330501, Qiagen) in Bio-Rad CFX384 PCR machine. Results were expressed as relative fold difference.
Wang J., Ghali S., Xu C., Mussatto C.C., Ortiz C., Lee E.C., Tran D.H., Jacobs J.P., Lagishetty V., Faull K.F., Moller T., Rossetti M., Chen X, & Koon H.W. (2018). Ceragenin CSA13 Reduces Clostridium difficile Infection in Mice by Modulating the Intestinal Microbiome and Metabolites. Gastroenterology, 154(6), 1737-1750.
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7

Systematic Gene Profiling of Cancer Pathways

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Systematic gene profiling was accomplished using Qiagen’s PCR array kit, according to the manufacturer’s protocol. Briefly, RNA from cell pellets were extracted and quantified. cDNA synthesis was performed using the RT2 First Strand Kit (Cat. No. 330401; Qiagen, GmbH), and subsequently combined with the RT2 SYBR® Green PCR master mix (Cat. No. 330504; Qiagen, GmbH). The master mix was then used in combination with the Human Cancer PathwayFinder RT2 lncRNA PCR Array (Cat. No. LAHS-002Z; Qiagen, GmbH), and the output data was analyzed using the GeneGlobe Data Analysis Center at http://www.qiagen.com/geneglobe.
Tang S.J., You G.R., Chang J.T, & Cheng A.J. (2021). Systematic Analysis and Identification of Dysregulated Panel lncRNAs Contributing to Poor Prognosis in Head-Neck Cancer. Frontiers in Oncology, 11, 731752.
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8

Investigating EMT and Stemness in 3D Cell Culture

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Cells were cultured 24 h over the matrigel or COL1-gel (0.5 μg/ml, rat tail, non pepsinized, Ibidi) supplemented with or without 2 ug/ml COL6A3 (MyBioSource). Cell culture medium was supplemented with or without 10 ng/ml OPN (R&D systems), 10 ng/ml IL-8 (Abcam). Total RNA was purified with Trizol (Invitrogen). cDNA was synthesized with RT2 First strand kit (Qiagen). Transcripts levels were measured by RT2Profiler™ PCR arrays, human stem cell and human epithelial to mesenchymal transition (EMT) using RT2 SYBR Green PCR master mix (Qiagen) and LightCycler480 (Roche). Fold expressions were calculated using the formula 2(−ΔΔCt), where ΔΔCt is ΔCt(sample)−ΔCt(control sample), ΔCt is Ct(gene of interest)−Ct(average from control gene setup) and Ct is the cycle at which the detection threshold is crossed.
Jokela T.A., Engelsen A.S., Rybicka A., Pelissier Vatter F.A., Garbe J.C., Miyano M., Tiron C., Ferariu D., Akslen L.A., Stampfer M.R., Lorens J.B, & LaBarge M.A. (2018). Microenvironment-Induced Non-sporadic Expression of the AXL and cKIT Receptors Are Related to Epithelial Plasticity and Drug Resistance. Frontiers in Cell and Developmental Biology, 6, 41.
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9

Quantitative PCR analysis of EMT and TGF-β/BMP pathways

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RNA (input of total RNA: 1.0 µg per reaction) was reverse transcribed using the RT2 First-Strand kit (Qiagen). Subsequently, profiler array-based quantitative PCR analysis was performed, following the instructions of the supplier, using the RT2 SYBR Green PCR Mastermix (Qiagen) and primer pre-coated RT2 Profiler PCR array plates (Qiagen) encompassing 84 pathway-focused genes per plate. Two different human-specific profiler PCR arrays were screened: pathways related to EMT signaling and the TGF-β/bone morphogenic protein (BMP), each on two independent biological replicates.
Soelch S., Beaufort N., Loessner D., Kotzsch M., Reuning U., Luther T., Kirchner T, & Magdolen V. (2021). Rab31-dependent regulation of transforming growth factor ß expression in breast cancer cells. Molecular Medicine, 27, 158.
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10

Inflammatory Cytokine and miRNA Expression

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Detection and quantification of gene expression was performed using a Mouse Inflammatory Cytokine and Receptors array (PAMM-011ZF; SABiosciences/QIAGEN, Frederick, USA) according to the manufacturer’s instructions. This PCR based array was selected as it includes 84 diverse genes important in the immune response, including genes encoding CC chemokines, CXC chemokines, interleukin cytokines, other cytokines, chemokine receptors, and cytokine receptors, as well as other genes involved in the inflammatory response. microRNA (miRNA) array was also performed using an Inflammatory Response miRNA PCR Array (MIMM-105Z; SABiosciences/QIAGEN) according to the manufacturer’s instructions. Real-time PCR amplification was performed using RT2 SYBR Green PCR Master Mix (QIAGEN). Primers were designed using web tool Primer 3 and synthesized by Eurofins MWG Operon (Huntsville, AL). Data analysis was performed using the ΔΔCT method according to the manufacturer’s protocol (SABiosciences).
Parmar T., Parmar V.M., Perusek L., Georges A., Takahashi M., Crabb J.W, & Maeda A. (2018). Lipocalin 2 plays an important role in regulating inflammation in retinal degeneration. Journal of immunology (Baltimore, Md. : 1950), 200(9), 3128-3141.
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