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Sybr green fastmix rox

Manufactured by Quanta Biosciences
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SYBR Green FastMix ROX is a ready-to-use solution for quantitative real-time PCR (qPCR) applications. It contains a proprietary hot-start DNA polymerase, SYBR Green I dye, and ROX passive reference dye, optimized for fast and sensitive detection of DNA targets.

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Lab products found in correlation

Quantitect primer assay, by Qiagen (5 mentions) Rneasy mini kit, by Qiagen (5 mentions) Superscript 3 first strand kit, by Thermo Fisher Scientific (5 mentions) 7300 real time instrument, by Thermo Fisher Scientific (4 mentions) Qubit 2.0 fluorometer, by Thermo Fisher Scientific (2 mentions) Agilent bioanalyzer tapestation 2200, by Agilent Technologies (1 mentions) Truseq rna kit, by Illumina (1 mentions) Rneasy lipid mini kit, by Qiagen (1 mentions) Taqman primer, by Thermo Fisher Scientific (1 mentions) High capacity cdna transcription kit, by Thermo Fisher Scientific (1 mentions) 7900ht fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Qiazol, by Qiagen (1 mentions) Gentlemacs m tube, by Miltenyi Biotec (1 mentions) Nextseq 500, by Illumina (1 mentions) Superscript 3 first strand synthesis kit, by Thermo Fisher Scientific (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)

Close spelling variants of this product

PerfeCTa SYBR Green FastMix ROX (79 citations) SYBR Green (48 citations) PerfeCTa SYBR Green FastMix Low ROX (30 citations) SYBR Green FastMix (27 citations) Perfecta SYBR Green FastMix ROX kit (4 citations) PerfeCTa SYBR Green QPCR FastMix ROX (2 citations)

6 protocols using sybr green fastmix rox

1

Quantitative Real-time PCR Analysis

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RNA was isolated with RNeasy Mini Kits (Qiagen). cDNA was generated with Superscript III First Strand kits (Invitrogen). Genes were measured by real time-RT PCR (qPCR) using SYBR Green FastMix ROX (Quanta Biosciences, Gaithersburg, MD) on a 7300 Real Time instrument (Applied Biosystems, CA). Expression was normalized to Gapdh. Primers were from Super Array Biosciences or QuantiTect Primer Assays (Qiagen).
Garg A.V., Amatya N., Chen K., Cruz J.A., Grover P., Whibley N., Conti H.R., Mir G.H., Sirakova T., Childs E.C., Smithgall T.E., Biswas P.S., Kolls J.K., McGeachy M.J., Kolattukudy P.E, & Gaffen S.L. (2015). MCPIP1 endoribonuclease activity negatively regulates interleukin-17-mediated signaling and inflammation. Immunity, 43(3), 475-487.
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2

RNA-Seq Analysis of Mouse and Human Cell Lines

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RNA-Seq mouse libraries were prepared with total RNA using a Qubit 2.0 fluorometer (Thermo Fisher) and Agilent Bioanalyzer TapeStation 2200 (Agilent Technologies). For OKF6-TERT2 in vitro infections, RNA-seq libraries (non-strand specific, paired-end) were prepared with the TruSeq RNA kit (Illumina). 100 nucleotides were determined from each end of cDNA fragments using the HiSeq platform. Single reads were aligned to the UCSC mouse or human reference genomes (mm10, GRCm38.75; Ensembl GRCh38) using TopHat2. Differential gene expression was assessed using DESeq (Bioconductor). Pairwise differential expression was quantified with Cuffdiff. Cufflinks was used to determine FPKM levels for each gene from the STAR alignment and was used as input for Cuffdiff. Read counts were then normalized across all samples and differentially expressed genes were determined by adjusted P-value with a threshold of 0.05.
For real-time PCR, RNA was isolated from tongue with RNeasy Mini Kits (QIAGEN), and cDNA generated with Superscript III First Strand Kits (Invitrogen). Real-time qPCR using SYBR Green FastMix ROX (Quanta Biosciences) was performed using a 7300 Real time instrument (Applied Biosystems) and normalized to GAPDH. Primers were from Superarray Biosciences or QuantiTect Primer Assays (QIAGEN).
Conti H.R., Bruno V.M., Childs E.E., Daugherty S., Hunter J.P., Mengesha B.G., Saevig D.L., Hendricks M.R., Coleman B.M., Brane L., Solis N., Cruz J.A., Verma A.H., Garg A.V., Hise A.G., Richardson J.P., Naglik J.R., Filler S.G., Kolls J.K., Sinha S, & Gaffen S.L. (2016). IL-17 receptor signaling in oral epithelial cells is critical for protection against oropharyngeal candidiasis. Cell host & microbe, 20(5), 606-617.
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3

RNA Isolation and qPCR Analysis

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Human biopsies were homogenized and RNA isolated with RNeasy Plus Mini kit (Qiagen). For qPCR, RNA was reverse transcribed using a High Capacity cDNA Transcription kit (Applied Biosystems). Quantitative qPCR was performed using the 7900HT Fast Real-Time PCR system (Applied Biosystems) with Taqman primers (Applied Biosystems). Expression was normalized to RPLP0. Mouse skin was homogenized in Qiazol (Qiagen), in GentleMACS M tubes (Miltenyi) and total RNA isolated using RNeasy Lipid Mini Kits (Qiagen). Cells were lysed in RLT buffer and RNA extraction was performed using RNeasy Mini Kit (Qiagen). cDNA was prepared using Superscript III First Strand kit (Invitrogen). qPCR was performed with QuantiTect Primer Assays (Qiagen) and SYBR Green Fast Mix ROX (Quanta Biosciences, Gaithersburg, MD). Expression was normalized to Gapdh for mouse skin and Rplp0 for keratinocytes. Samples were analyzed on a 7300 Real Time instrument (Applied Biosystems, CA). Ct values were obtained for target and housekeeping genes and ΔCt values calculated. Expression was normalized to Gapdh for mouse skin and Rplp0 for keratinocytes. Gene induction was then transformed to a linear scale via calculation of 2− ΔCt (relative expression).
Monin L., Gudjonsson J.E., Childs E.E., Amatya N., Xing X., Verma A.H., Coleman B.M., Garg A., Killeen M., Mathers A., Ward N.L, & Gaffen S.L. (2016). MCPIP1/Regnase-1 restricts IL-17A- and IL-17C-dependent skin inflammation. Journal of immunology (Baltimore, Md. : 1950), 198(2), 767-775.
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4

Characterization of IMP2-Bound Transcripts

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RNA was isolated with RNeasy Mini Kits (Qiagen), and cDNA was synthesized by SuperScript III First Strand Kits (Thermo Fisher Scientific, Waltham MA). Real-time qPCR was performed with SYBR Green FastMix ROX (Quanta Biosciences) on a 7300 Real-Time instrument (Applied Biosystems), normalized to Gapdh. Primers were from QuantiTect Primer Assays (QIAGEN). RNASeq libraries were prepared from MEF mRNA (Nextera XT Kit). RNASeq was performed on Illumina NextSeq 500 by the Health Sciences Sequencing Core at the University of Pittsburgh.
For RIP-Seq, libraries were generated using SMART-Seq v4 Ultra Low Input RNA kit (Takara Biosciences) and sequenced on a NextSeq500. Raw reads were aligned to mm10 using STAR (119 (link)) followed by identification of global binding of IMP2 using Piranha (55 (link)). Differentially enriched IMP2 interactions were identified with DESeq2 (56 (link)) by comparing 3 replicates of IMP2-RIP-seq data and control IgG. Identification of IMP2 motifs (35 (link), 120 (link)) was performed using MDS2 (57 (link)) and viewed using Integrative Genomics Viewer (Broad Institute).
Bechara R., Amatya N., Bailey R.D., Li Y., Aggor F.E., Li D.D., Jawale C.V., Coleman B.M., Dai N., Gokhale N.S., Taylor T.C., Horner S.M., Poholek A.C., Bansal A., Biswas P.S, & Gaffen S.L. (2021). The m6A reader IMP2 directs autoimmune inflammation through an IL-17- and TNFα-dependent C/EBP transcription factor axis. Science immunology, 6(61), eabd1287.
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5

RNA Isolation and qPCR Gene Expression Analysis

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Total RNA was isolated using RNeasy Mini Kits (Qiagen). cDNA synthesis was performed using Superscript III First Strand kits (Invitrogen, Carlsbad CA). Genes were measured by real time-reverse transcriptase PCR (qPCR) using SYBR Green FastMix ROX from Quanta Biosciences (Gaithersburg, MD). PCR reactions were performed on a 7300 Real Time PCR Systems instrument (Applied Biosystems, CA). Expression was normalized to Gapdh. Primers were from Super Array Biosciences or QuantiTect Primer Assays (Qiagen).
Conti H.R., Whibley N., Coleman B.M., Garg A.V., Jaycox J.R, & Gaffen S.L. (2015). Signaling through IL-17C/IL-17RE Is Dispensable for Immunity to Systemic, Oral and Cutaneous Candidiasis. PLoS ONE, 10(4), e0122807.
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6

Antioxidant Gene Expression Analysis in Bovine Cells

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These procedures have been thoroughly described by Bionaz and Loor (2008, 2011) . Briefly, RNA quality was measured using an Agilent 2100 Bioanalyzer (Agilent, Santa Clara, CA). All samples had an RNA integrity number greater than 7.3. Total RNA was reverse-transcribed using the SuperScript III First Strand Synthesis kit (Thermo Fisher Scientific, Waltham MA). Realtime reverse-transcription PCR (quantitative; qPCR) analysis was performed using SYBR Green Fast Mix ROX (Quanta Biosciences, Gaithersburg, MD) with a 6-point standard curve. A comprehensive literature search (Chorley et al., 2012; (link)Niture and Jaiswal, 2012; (link)Jin et al., 2016a) was conducted to select NFE2L2 target genes and regulators with key roles in the antioxidant mechanisms evaluated (Supplemental Table S1; https:// doi .org/ 10 .3186/ jds .2017 -14257). This included the cell apoptosis regulator BCL2, the glutathione metabolism system (GCLC, GCLM, GPX1, GSR, GSTM1, ME1 and TALDO1), the thioredoxin enzyme system (TXNRD1, TXN), the heme and Fe metabolism system (HMOX1, FECH, FTH1, PIR), and NAD (P) H quinone dehydrogenase 1 gene (NQO1), KEAP-dependent regulators (NFE2L2, KEAP1, CUL3) and KEAPindependent regulators (NFKB1, MAPK8, MAPK12, MAPK14, RXRA). Gene symbols, primer features, and qPCR performance are reported in Supplemental Table S2 (https:// doi .org/ 10 .3186/ jds .2017 -14257).
Han L.Q., Zhou Z., Ma Y., Batistel F., Osorio J.S, & Loor J.J. (2018). Phosphorylation of nuclear factor erythroid 2-like 2 (NFE2L2) in mammary tissue of Holstein cows during the periparturient period is associated with mRNA abundance of antioxidant gene networks. Journal of dairy science, 101(7).
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