Takyon sybr green pcr kit

Manufactured by Eurogentec

Sourced in France

The Takyon SYBR Green PCR kit is a reagent kit designed for real-time quantitative PCR (qPCR) analysis. It contains the necessary components, including SYBR Green I dye, for performing real-time PCR reactions. The kit is intended to be used with compatible real-time PCR instruments.

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Lab products found in correlation

Rneasy mini kit, by Qiagen (5 mentions) Steponeplus apparatus, by Thermo Fisher Scientific (4 mentions) Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (3 mentions) Quantitect reverse transcription kit, by Qiagen (1 mentions) Luna universal one step rt qpcr kit, by New England Biolabs (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Abi7300 apparatus, by Thermo Fisher Scientific (1 mentions) Iscript reverse transcriptase, by Bio-Rad (1 mentions)

6 protocols using takyon sybr green pcr kit

Quantitative RT-PCR Analysis of Colon Samples

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According to the manufacturer’s instructions, total RNA was isolated from colon samples or cell suspensions using an RNeasy Mini Kit (Qiagen) Quantitative RT-PCR was performed using QuantiTect Reverse Transcription Kit (Qiagen) and then a Takyon SYBR Green PCR kit (Eurogentec) or Luna Universal One-Step RT-qPCR Kit (New England Biolabs) in a StepOnePlus apparatus (Applied Biosystems) with specific mouse oligonucleotides.

Michaudel C., Danne C., Agus A., Magniez A., Aucouturier A., Spatz M., Lefevre A., Kirchgesner J., Rolhion N., Wang Y., Lavelle A., Galbert C., Da Costa G., Poirier M., Lapière A., Planchais J., Nádvorník P., Illes P., Oeuvray C., Creusot L., Michel M.L., Benech N., Bourrier A., Nion-Larmurier I., Landman C., Richard M.L., Emond P., Seksik P., Beaugerie L., Arguello R.R., Moulin D., Mani S., Dvorák Z., Bermúdez-Humarán L.G., Langella P, & Sokol H. (2022). Rewiring the altered tryptophan metabolism as a novel therapeutic strategy in inflammatory bowel diseases. Gut, 72(7), 1296-1307.

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Quantitative RT-PCR analysis of gene expression

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Total RNA was isolated from colon samples using an RNeasy Mini Kit (Qiagen, Hilden, Germany), according to the manufacturer’s instructions. Quantitative RT–PCR was performed using SuperScript II Reverse Transcriptase (Life Technologies, Saint Aubin, France) and then a Takyon SYBR Green PCR kit (Eurogentec, Liège, Belgium) in a StepOnePlus apparatus (Applied Biosystems, Foster City, CA, USA) with specific mouse oligonucleotides. The oligonucleotides used were as follows: Gapdh (sense) 5′-AACTTTGGCATTGTGGAAGG-3′ and (antisense) 5′-ACACATTGGGGGTAGGAACA-3′; Il17a (sense) 5′-TTTAACTCCCTTGGCGCAAAA-3′ and (antisense) 5′-CTTTCCCTCCGCATTGACAC-3′; Il22 (sense) 5′-CATGCAGGAGGTGGTACCTT-3′ and (antisense) 5′-CAGACGCAAGCATTTCTCAG-3′; Reg3g (sense) 5′-TTCCTGTCCTCCATGATCAAAA-3′ and (antisense) 5′-CATCCACCTCTGTTGGGTTCA-3′; and Reg3b (sense) 5′-ATGCTGCTCTCCTGCCTGATG-3′ and (antisense) 5′-CTAATGCGTGCGGAGGGTATATTC-3′. We used the 2^−ΔΔ^Ct quantification method with mouse Gapdh as an endogenous control and the WT or WT→GF group as a calibrator.

Lamas B., Richard M.L., Leducq V., Pham H.P., Michel M.L., Da Costa G., Bridonneau C., Jegou S., Hoffmann T.W., Natividad J.M., Brot L., Taleb S., Couturier-Maillard A., Nion-Larmurier I., Merabtene F., Seksik P., Bourrier A., Cosnes J., Ryffel B., Beaugerie L., Launay J.M., Langella P., Xavier R.J, & Sokol H. (2016). CARD9 impacts colitis by altering gut microbiota metabolism of tryptophan into aryl hydrocarbon receptor ligands. Nature medicine, 22(6), 598-605.

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Quantification of Gut Microbiota Composition

Cited 2 times

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Fecal, cecal and small intestinal content genomic DNA was extracted from the weighted stool samples using a method that was previously described^{31 (link)}, which is based on the Godon DNA extraction method. Quantifications of all bacteria and B. wadsworthia were performed by qPCR using TaqMan Gene Expression Assays (Life technologies) and Takyon SYBR Green PCR kit (Eurogentec). All bacteria was quantified using the following oligonucleotides: (sense) 5′-CGGTGAATACGTTCCCGG-3′ and (antisense) 5′-TACGGCTACCTTGTTACGACTT-3′ and (probe) 5′-CTTGTACACACCGCCCGTC-3′. B. wadsworthia was quantified using specific primers for the tpa gene (accession no. AF269146): (sense) 5′-CGCCGGTATCGAAATCGTGA-3′ and (antisense) 5′-ATTCGCGGAAGGAGCGAGAG-3′. Sulfite-reducing bacteria were quantified using specific primers for the dsra gene (encoding a dissimilatory sulfite reductase alpha subunit) as described by Devkota^{5 (link)}.

Natividad J.M., Lamas B., Pham H.P., Michel M.L., Rainteau D., Bridonneau C., da Costa G., van Hylckama Vlieg J., Sovran B., Chamignon C., Planchais J., Richard M.L., Langella P., Veiga P, & Sokol H. (2018). Bilophila wadsworthia aggravates high fat diet induced metabolic dysfunctions in mice. Nature Communications, 9, 2802.

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Colon RNA Extraction and qRT-PCR

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Total RNA was isolated from colon samples or cell suspensions using RNeasy Mini Kit (Qiagen) and quantitative RT-PCR performed using SuperScript II Reverse Transcriptase (Life Technologies) and then a Takyon SYBR Green PCR kit (Eurogentec) in a StepOnePlus apparatus (Applied Biosystems) with specific mouse oligonucleotides (Supplemental Table S1). We used the 2 − ΔΔCt quantification method with mouse Gapdh as an endogenous control and the GF WT or WF → GF WT or WT → GF Card9^−/− group as a calibrator.

Danne C., Lamas B., Lavelle A., Michel M.L., Da Costa G., Pham H.P., Lefevre A., Bridonneau C., Bredon M., Planchais J., Straube M., Emond P., Langella P, & Sokol H. (2024). Dissecting the respective roles of microbiota and host genetics in the susceptibility of Card9−/− mice to colitis. Microbiome, 12, 76.

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Quantitative RT-PCR Analysis of Colon RNA

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Total RNA was isolated from colon samples using RNeasy Mini Kit (Qiagen). Quantitative RT-PCR was performed using iScript Reverse Transcriptase (Bio-Rad) and then a Takyon SYBR Green PCR kit (Eurogentec) in an ABI 7300 apparatus (Applied Biosystems) with specific mouse oligonucleotides (Eurogentec) described in Table S3. Thermal cycle conditions were 10 min at 95°C, then 40 cycles of 15 s at 94°C, 45 s at 58°C, and 30 s at 72°C followed by 5 min at 72°C. We used the

2 begin superscript negative uppercase delta uppercase delta cycle threshold end superscript

quantification method with mouse Rpl19 as an endogenous control and the OVA-tolerized mice not exposed to

food-grade silicon dioxide

or nonobese diabetic (NOD) mice expressing HLA-DQ8 (NOD/DQ8) not exposed to

food-grade silicon dioxide

group as calibrator.

Lamas B., Martins Breyner N., Malaisé Y., Wulczynski M., Galipeau H.J., Gaultier E., Cartier C., Verdu E.F, & Houdeau E. (2024). Evaluating the Effects of Chronic Oral Exposure to the Food Additive Silicon Dioxide on Oral Tolerance Induction and Food Sensitivities in Mice. Environmental Health Perspectives, 132(2), 027007.

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Quantitative RT-PCR Analysis of Colon Samples

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Total RNA was isolated from colon samples using an RNeasy Mini Kit (Qiagen, Hilden, Germany), including a DNAse treatment step, according to the manufacturer’s instructions. Quantitative RT-PCR was performed using SuperScript II Reverse Transcriptase (Life Technologies, Saint Aubin, France) followed by a Takyon SYBR Green PCR kit (Eurogentec, Liège, Belgium) in a StepOnePlus apparatus (Applied Biosystems, Foster City, CA, USA) with specific mouse oligonucleotides. The oligonucleotides used were as follows: GAPDH—sense: 5′-AACTTTGGCATTGTGGAAGG-3′; antisense: 5′-ACACATTGGGGGTAGGAACA-3′, Reg3g—sense: 5′-TTCCTGTCCTCCATGATCAAAA-3′; antisense: 5′-CATCCACCTCTGTTGGGTTCA-3′. We used the 2^−ΔΔCt quantification method with mouse GAPDH as an endogenous control and calibrated the assay to the wild type.

Sovran B., Planchais J., Jegou S., Straube M., Lamas B., Natividad J.M., Agus A., Dupraz L., Glodt J., Da Costa G., Michel M.L., Langella P., Richard M.L, & Sokol H. (2018). Enterobacteriaceae are essential for the modulation of colitis severity by fungi. Microbiome, 6, 152.

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