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Superscript 3 reverse transcriptase

Manufactured by Qiagen
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Sourced in Germany, United States

SuperScript III Reverse Transcriptase is a laboratory equipment product used for the conversion of RNA to complementary DNA (cDNA) in reverse transcription reactions. It is a thermostable, RNase H-deficient reverse transcriptase enzyme that can be used to generate high-quality cDNA from a variety of RNA templates.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (3 mentions) Rneasy mini kit, by Qiagen (3 mentions) Thermal cycler dice real time system, by Takara Bio (2 mentions) 2 super array pcr master mix, by Qiagen (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) Lightcycler 480 detection system, by Roche (2 mentions) Sybr green mix, by Thermo Fisher Scientific (2 mentions) Abi prism 7000 analyzer, by Thermo Fisher Scientific (2 mentions) Sybr green master mix, by Thermo Fisher Scientific (1 mentions) Oligo dt, by Thermo Fisher Scientific (1 mentions) 7500 fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Tnf mm00443258 m1, by Thermo Fisher Scientific (1 mentions) Tbx21 mm00450960 m1, by Thermo Fisher Scientific (1 mentions) Sv total rna isolation kit, by Promega (1 mentions) Quantitect sybr green pcr kit, by Qiagen (1 mentions) Rneasy kit, by Qiagen (1 mentions) Viia 7 real time pcr system, by Thermo Fisher Scientific (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) High capacity cdna reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Cfx96 detection system, by Bio-Rad (1 mentions)

Spelling variants of this product

Reverse transcriptase (12 citations) SuperScript III (9 citations)

12 protocols using superscript 3 reverse transcriptase

1

Retinal microRNA expression analysis

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Total RNA was extracted from the retinal tissues of the normoxia, OIR, and OIR-treated groups using TRIzol® reagent (Invitrogen; Thermo Fisher Scientific, Inc.) and cleaned with the RNeasy_MinElute Cleanup kit (Qiagen GmbH, Hilden, Germany). SuperScript III Reverse Transcriptase (Qiagen GmbH) was used to reverse transcribe the total RNA, and cDNA was amplified by polymerase chain reaction using the 2_Super Array PCR Master mix (Qiagen GmbH). The MicroRNA PCR array (SuperArray Bioscience Corporation, Frederick, MD, USA) was performed in a Thermal Cycler Dice Real-Time system (Takara Bio, Inc.) according to the manufacturer's protocol. The results were normalized to U6B levels using the 2−ΔΔCq method (21 (link)).
Zhang W., Chen L., Geng J., Liu L, & Xu L. (2019). β-elemene inhibits oxygen-induced retinal neovascularization via promoting miR-27a and reducing VEGF expression. Molecular Medicine Reports, 19(3), 2307-2316.
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2

Quantitative PCR analysis of thymic stromal cells

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RNA from sorted thymic stromal cell populations was purified using the Absolutely RNA Microprep kit (Stratagene). RNA from stromal fraction of the thymus, whole thymus, spleen, and skin was prepared using TRIzol (Invitrogen). RNA was reverse-transcribed into cDNA using oligo-dT (Invitrogen) and Superscript III reverse transcriptase (Qiagen). Ifi30 (GILT) transcript levels were assessed by real-time PCR using the exon 1–2 amplifying primers 5’-AGGCACAACCACCTGCAA-3’ and 5’-AGCGACAAGCTCCACACA-3’, and Tyrp1 was detected using the exon 6–7 amplifying primers 5’-CACTTTCACTGATGCGGTCTTTGAC-3’ and 5’-TGTCCAATAGGTGCGTTTTCCAAC-3’. Products were detected using SYBR green master mix (Applied Biosystems). Sample quantities were normalized to cyclophilin signal (SuperArray Biosciences). Real-time PCR reactions were run on a 7500 Fast Real-Time PCR System (Applied Biosystems), and data analyzed according to the standard curve method using Excel (Microsoft).
Rausch M.P., Meador L.R., Metzger T.C., Li H., Qiu S., Anderson M.S, & Hastings K.T. (2020). GILT in thymic epithelial cells facilitates central CD4 T cell tolerance to a tissue-restricted, melanoma-associated self antigen. Journal of immunology (Baltimore, Md. : 1950), 204(11), 2877-2886.
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3

Quantitative PCR Analysis of Mouse Gastric Immune Markers

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RNA was isolated from scraped gastric mucosa or from FACS-sorted cells using the RNeasy Mini kit (QIAGEN) according to the manufacturer's instructions. Complementary DNA synthesis was performed using Superscript III reverse transcriptase (QIAGEN). Quantitative PCR reactions for the candidate genes were performed using TaqMan gene expression assays (Csf2 Mm01290062_m1, Cxcl9 Mm00434946_m1, Cxcl10 Mm00445235_m1, Cxcl11 Mm00444662_m1, Foxp3 Mm00475162_m1, Gzmb Mm00442837_m1, Hprt Mm03024075_m1, Ifng Mm01168134_m1, Il2 Mm00434256_m1, Il4 Mm00445259_m1, Il10 Mm01288386_m1, Il12a Mm00434169_m1, Il13 Mm00434204_m1, Il23 Mm00518984_m1, Tbx21 Mm00450960_m1, Tnf Mm00443258_m1, Rorc Mm01261022_m1; Applied Biosystems). Complementary DNA samples were analyzed in duplicate using a Light Cycler 480 detection system (Roche) and gene expression levels for each sample were normalized to HPRT expression. Mean relative gene expression was determined, and the differences were calculated using the 2ΔC(t) method.
Arnold I.C., Zhang X., Artola-Boran M., Fallegger A., Sander P., Johansen P, & Müller A. (2019). BATF3-dependent dendritic cells drive both effector and regulatory T-cell responses in bacterially infected tissues. PLoS Pathogens, 15(6), e1007866.
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4

Quantitative PCR Analysis of T Cell RNAs

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Total RNAs were isolated from T cells with Trizol reagent (Invitrogen). A total of 0.5 μg of RNA was converted to cDNA using SuperScript III Reverse Transcriptase (Qiagen) with random hexamer primers. qPCR was performed using the ABI Prism 7000 Analyzer (Applied Biosystems) with SYBR Green mix (Applied Biosystems). All primers are listed in Supplementary Table 2.
Li Q., Zou J., Wang M., Ding X., Chepelev I., Zhou X., Zhao W., Wei G., Cui J., Zhao K., Wang H.Y, & Wang R.F. (2014). Critical role of histone demethylase Jmjd3 in the regulation of CD4+ T cell differentiation. Nature communications, 5, 5780.
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5

Quantitative PCR Analysis of Immune Genes

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RNA was isolated from FACS-sorted cells using an RNeasy minikit (Qiagen) according to the manufacturer's instructions. cDNA synthesis was performed using SuperScript III reverse transcriptase (Qiagen). Quantitative PCRs for the candidate genes were performed using TaqMan gene expression assays as follows: for Hprt, Mm03024075_m1; for Tgfb1, Mm01178820_m1; for IL-10, Mm01288386_m1; for IL-23, Mm00518984_m1; for Il6, Mm00446190_m1; for Ptgs2, Mm00478374_m1; for Tnf, Mm00443258_m1; for IL-12b, Mm01288989_m1) (all from Applied Biosystems by Thermo Fisher Scientific). cDNA samples were analyzed in duplicate using a Light Cycler 480 detection system (Roche), and gene expression levels were normalized to hypoxanthine phosphoribosyltransferase (HPRT) expression for each sample. Mean relative gene expression levels were determined, and the differences were calculated using the threshold cycle (2ΔCT) method.
Altobelli A., Bauer M., Velez K., Cover T.L, & Müller A. (2019). Helicobacter pylori VacA Targets Myeloid Cells in the Gastric Lamina Propria To Promote Peripherally Induced Regulatory T-Cell Differentiation and Persistent Infection. mBio, 10(2), e00261-19.
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6

Quantitative Real-Time RT-PCR Analysis

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Total RNA was prepared from TA muscles using Promega SV Total RNA Isolation kit. Complementary DNA (cDNA) generated with Invitrogen SuperScript III Reverse Transcriptase was analysed by quantitative real-time RT–PCR using Qiagen QuantiTect SYBR Green PCR Kit. All data were normalized to GAPDH and actin expression. The oligonucleotide primers used are shown in Supplementary Table 1.
Milan G., Romanello V., Pescatore F., Armani A., Paik J.H., Frasson L., Seydel A., Zhao J., Abraham R., Goldberg A.L., Blaauw B., DePinho R.A, & Sandri M. (2015). Regulation of autophagy and the ubiquitin–proteasome system by the FoxO transcriptional network during muscle atrophy. Nature Communications, 6, 6670.
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7

qPCR Analysis of Inflammatory Markers

Cited 1 time
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RNA was isolated with RNeasy kit (QIAGEN, Hilden, Germany) and reverse transcribed to cDNA with Super-Script III reverse transcriptase (QIAGEN). The cDNA was used as template for real time (RT)-PCR using SYBER green PCR Master Mix with primers for ICAM-1, CCL2, TNF-α, IL-1β, IL-6, CXCL1, NOS2, and P2X7.9 (link) Gene expression was assessed using a StepOne Real Time PCR system (ABI 7900 Sequence Detection System). The cDNA samples were run in triplicate. Samples were normalized according to the content of 18S rRNA.
Yu J.S., Daw J., Portillo J.A, & Subauste C.S. (2021). CD40 Expressed in Endothelial Cells Promotes Upregulation of ICAM-1 But Not Pro-Inflammatory Cytokines, NOS2 and P2X7 in the Diabetic Retina. Investigative Ophthalmology & Visual Science, 62(12), 22.
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8

Quantitative PCR Analysis of T Cell RNAs

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Total RNAs were isolated from T cells with Trizol reagent (Invitrogen). A total of 0.5 μg of RNA was converted to cDNA using SuperScript III Reverse Transcriptase (Qiagen) with random hexamer primers. qPCR was performed using the ABI Prism 7000 Analyzer (Applied Biosystems) with SYBR Green mix (Applied Biosystems). All primers are listed in Supplementary Table 2.
Li Q., Zou J., Wang M., Ding X., Chepelev I., Zhou X., Zhao W., Wei G., Cui J., Zhao K., Wang H.Y, & Wang R.F. (2014). Critical role of histone demethylase Jmjd3 in the regulation of CD4+ T cell differentiation. Nature communications, 5, 5780.
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9

Quantifying Gene Expression in Intestinal Tissues

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RNA was isolated from snap-frozen colonic tissue (combined proximal, mid and distal colon sections), isolated lamina propria cells, BMDM or from FACS-sorted cells, using the RNeasy Mini kit (QIAGEN) according to manufacturer’s instructions, including an on-column DNase I digestion step. cDNA synthesis was performed using Superscript III reverse transcriptase (QIAGEN) or High Capacity cDNA Reverse Transcriptase (Applied Biosystems, Life Technologies). Quantitative PCR (qPCR) reactions for the candidate genes were performed using TaqMan gene expression assays (Life Technologies). cDNA samples were analysed in duplicate using the CFX96 detection system (Bio-Rad Laboratories) or ViiA™ 7 Real Time PCR System (Life Technologies), and gene expression levels for each sample were normalized to HPRT. Mean relative gene expression was determined, and the differences were calculated using the 2ΔC(t) method. Primer pairs and probes: TaqMan gene expression assays for mouse Hprt (Mm01545399_m1), Il23a (Mm00518984_m1) and (Mm01160011_g1).
Arnold I.C., Mathisen S., Schulthess J., Danne C., Hegazy A.N, & Powrie F. (2015). CD11c+ monocyte/macrophages promote chronic Helicobacter hepaticus-induced intestinal inflammation through the production of IL-23. Mucosal Immunology, 9(2), 352-363.
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10

Quantifying Tumor Metastasis Genes

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Forty-eight hours after siRNA knockdown, RNA was extracted from the cells using Trizol (Invitrogen) and cleaned with the RNeasy_MinEluteTM Cleanup Kit (Qiagen, Valencia, CA). Subsequently, total RNA was reverse transcribed using SuperScript III Reverse Transcriptase (Qiagen), and complementary DNA was amplified by the polymerase chain reaction (PCR) using 2_Super Array PCR master mix (Qiagen). Real-time PCR was then performed on each sample with the Human Tumor Metastasis RT2 ProfilerTM PCR Array (SuperArray Bioscience) in a Thermal Cycler Dice Real Time System (Takara TP800, Japan) according to the manufacturer’s instructions. Data were normalized to GAPDH levels by the 2-ΔΔCt method.
Wu G., Liu H., He H., Wang Y., Lu X., Yu Y., Xia S., Meng X, & Liu Y. (2014). miR-372 down-regulates the oncogene ATAD2 to influence hepatocellular carcinoma proliferation and metastasis. BMC Cancer, 14, 107.
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