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Superscript 2

Manufactured by Roche
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Sourced in France, China, Canada

Superscript II is a reverse transcriptase enzyme used in the process of converting RNA to complementary DNA (cDNA) for various molecular biology applications. It provides efficient and reliable cDNA synthesis.

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Lab products found in correlation

Quantitect sybr green rt pcr kit, by Qiagen (3 mentions) Trizol, by Thermo Fisher Scientific (3 mentions) Dna engine opticon, by Bio-Rad (2 mentions) Trizol, by Merck Group (2 mentions) Lightcycler 480 2, by Roche (1 mentions) Lightcycler, by Roche (1 mentions) Rnase free dnase 1, by Qiagen (1 mentions) Rneasy kit, by Qiagen (1 mentions) Qiashredder column, by Qiagen (1 mentions) Sybr green, by Roche (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Sybr green master mix, by Thermo Fisher Scientific (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Nucleospin rna kit, by Macherey-Nagel (1 mentions) Random hexamer primer, by Roche (1 mentions) Sybr green kit, by Roche (1 mentions) Sybr green assay, by Thermo Fisher Scientific (1 mentions) Random hexamer, by Roche (1 mentions) Dnase treatment, by Qiagen (1 mentions) Cfx system, by Bio-Rad (1 mentions)

Spelling variants of this product

Superscript II reverse transcriptase (7 citations) Superscript II Reverse Transcription (3 citations) Superscript II RT enzyme (3 citations) SuperScript Reverse Transcriptase type II (2 citations)

12 protocols using superscript 2

1

Quantitative RT-PCR for Gene Expression Analysis

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RNA was reverse transcribed with SuperScript II (Roche). Real-time quantitative PCR was performed using the QuantiTect SYBR Green RT-PCR kit (Qiagen) on the Light Cycler 480 II (Roche). Gene expression was quantitated relative to β-actin and then normalized to the corresponding WT entheses of the same age isolated on the same day (for enthesis RNA) or control-treated chondrocytes (for primary chondrocyte RNA) for each sample, using the methods of Livak and Schmittgen (49 (link)).
Rana R., Baker J.T., Sorsby M., Jagga S., Venkat S., Almardini S, & Liu E.S. (2023). Impaired 1,25-dihydroxyvitamin D3 action underlies enthesopathy development in the Hyp mouse model of X-linked hypophosphatemia. JCI Insight, 8(17), e163259.
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2

Quantifying CTGF and CYR61 mRNA in HEK293A

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HEK293A cell lines (WT, LATS1/2KO and PTPN14KO) were grown in one 60mm dish at 50% confluence. Cells were detached by trypsinization and lysed using QIAshredder columns (Qiagen). Total RNA was extracted using RNeasy kit (Qiagen) and DNA was degraded using RNase-free DNase I (Qiagen) following manufacturer instructions. RNA was reverse transcribed into cDNA using random hexanucleotides (Microsynth) and SuperScript II polymerase (Roche). The relative abundance of CTGF and CYR61 mRNA was determined using a Roche LightCycler and SYBRgreen (Roche). GAPDH was used as a reference gene.
The following oligos were used: one full FTMS scan (350-1600 m/z) at 120'000 resolution followed by fifteen MS/MS scans in the Ion Trap. Charge states lower than two and higher than seven were rejected.
Selected ions were isolated using a quadrupole mass filter of 2.0 m/z isolation window.
Precursors with MS signal that exceeded a threshold of 500 were fragmented (CID, Normalized Collision Energy 35%). Selected ions were dynamical excluded for 30 s.
Uliana F., Ciuffa R., Mishra R., Fossati A., Frommelt F., Mehnert M., Birkeland E.S., Peter M., Tapon N., Aebersold R., & Gstaiger M. (2022). Systematic dissection of phosphorylation-dependent YAP1 complex formation elucidates a key role for PTPN14 in Hippo signal integration.
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3

Quantifying Periarticular Chondrocyte PTHrP mRNA

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Humeri were dissected free of muscle and connective tissue and following removal of both growth plates, marrow was flushed with phosphate-buffered saline to permit isolation of cortical bone, which was then homogenized in Trizol (Thermo Fisher Scientific). Total RNA was precipitated using 100% ethanol and purified using the RNeasy mini kit (Qiagen). To evaluate PTHrP mRNA expression the head of the humerus was embedded vertically in OCT. Sequential cryosections were stained for 30 seconds in Safranin-O to allow isolation of the peri-articular chondrocytes. Chondrocyte RNA was isolated from the first 50 microns using the RNeasy mini kit (Qiagen). In initial studies, humeri were then reembedded horizontally and cryosectioned to confirm that only sections with periarticular chondrocytes had been included in the RNA isolation. RNA was reverse transcribed with SuperScript™ II (Roche). Quantitative real time-PCR was performed using the QuantiTect SYBR Green RT-PCR kit (Qiagen) on an Opticon DNA engine (MJ Research). Gene expression was normalized to that of a control gene for each sample, using the methods of Livak and Schmittgen.(5 (link))
Liu E.S., Martins J.S., Raimann A., Chae B.T., Brooks D.J., Jorgetti V., Bouxsein M.L, & Demay M.B. (2016). 1,25-dihydroxyvitamin D alone improves Skeletal Growth, Microarchitecture and Strength in a Murine model of XLH, despite enhanced FGF23 expression. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 31(5), 929-939.
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4

Quantitative RNA Profiling from Cell Samples

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RNeasy Plus Mini Kit (Qiagen, Germantown, MD) was used to extract total RNA. First-strand cDNA was synthesized from 2 µg of total RNA using Superscript II reverse transcriptase (Roche, Mannheim, Germany). qPCR was performed in a 20 µL reaction mixture containing 10 µL of SYBR Green Master Mix (Applied Biosystems), 5 µL of 1 mM primers, and 100 ng of cDNA template using a Bio-Rad CFT Connect thermocycler. Data were normalized to glyceraldehyde 3-phosphate dehydrogenase. IDT synthesized primers used in qPCR can be found in Table 1.
Khalil M.I., Yang C., Vu L., Chadha S., Nabors H., James C.D., Morgan I.M, & Pyeon D. (2024). The membrane-associated ubiquitin ligase MARCHF8 stabilizes the human papillomavirus oncoprotein E7 by degrading CUL1 and UBE2L3 in head and neck cancer. Journal of Virology, 98(2), e01726-23.
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5

Quantification of Hepatitis Delta Virus

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For HDV quantification, total intracellular RNA was extracted using the Nucleospin RNA Kit (Macherey-Nagel AG). cDNA was synthesized from 100 ng total RNA using Superscript II and random hexamer primers (Roche Diagnosis). Quantification of HDV was performed by quantitative reverse-transcription PCR as described,19 (link) using the primers described by Scholtes & colleagues20 (link) and EEF1A1 as a housekeeping gene.
Tseligka E.D., Conzelmann S., Cambet Y., Schaer T., Negro F, & Clément S. (2022). Identification of selective hepatitis delta virus ribozyme inhibitors by high-throughput screening of small molecule libraries. JHEP Reports, 5(3), 100652.
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6

Quantitative Gene Expression Analysis

Cited 2 times
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Gene transcription analyses were performed by qRT-PCR after total RNA extraction with Trizol (Sigma Aldrich), reverse transcription using Superscript II on 500 μg total mRNA, and quantified with SYBRGreen kit (Roche Diagnosis, Meylan, France) as delta Ct (cycle threshold) as previously described [1 (link),2 (link)]. Results were normalized to hypoxanthine guanine phosphoribosyl transferase (HPRT) standard gene quantification and performed in three to four biological replicates. The list of primers has been previously published [12 (link),23 (link)].
Berger E, & Géloën A. (2023). FABP4 Controls Fat Mass Expandability (Adipocyte Size and Number) through Inhibition of CD36/SR-B2 Signalling. International Journal of Molecular Sciences, 24(2), 1032.
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7

Gene Expression Analysis in Colon and Pancreas

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RNA was extracted from colon samples with TRIzol (Sigma). RNA from pancreas samples was extracted with GTC solution, following a standard phenol–chloroform protocol (reagents from Sigma), followed by DNAse treatment (Qiagen). cDNA was synthesized using Random Hexamers (Roche) and SuperScript II reverse transcriptase. mRNA was quantified by SYBR green assay (Applied Biosystems), with normalization to GAPDH. The following primers were used: human-AID (forward) 5′-AAA TGT CCG CTG GGC TAA GG-3′, (reverse) 5′-GGA GGA AGA GCA ATT CCA CGT-3′; human-GAPDH (forward) 5′-GAA GGT GAA GGT CGG AGT C-3′, (reverse) 5′-GAA GAT GGT GAT GGG ATT TC-3′; mouse AID (forward) 5′-ACC TTC GCA ACA AGT CTG GCT-3′, (reverse) 5′-AGC CTT GCG GTC TTC ACA GAA-3′; mouse-GAPDH (forward) 5′-TGA AGC AGG CAT CTG AGG G-3′, (reverse) 5′-CGA AGG TGG AAG AGT GGG AG-3′; mouse-TNF-α (forward) 5′-AGC CCA CGT CGT AGC AAA CCA-3′, (reverse) 5′-ACA ACC CA CGG CTG GCA CC-3′.
Pérez-García A., Pérez-Durán P., Wossning T., Sernandez I.V., Mur S.M., Cañamero M., Real F.X, & Ramiro A.R. (2015). AID-expressing epithelium is protected from oncogenic transformation by an NKG2D surveillance pathway. EMBO Molecular Medicine, 7(10), 1327-1336.
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8

Cardiac Gene Expression Analysis in Rat LV

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Total RNA isolation from rat LV (non-infarcted LV myocardium surrounding the infarcted area) specimens with TRIzol (Invitrogen) as directed by the manufacturer. Reverse transcription was carried out with oligo (dT) primers and Superscript II (Roche). Next, the mRNA amounts of cardiac genes were assessed by qRT-PCR on a Bio-Rad CFX system. Table 1 shows the primers and probes for IL-6, IL-1β, IL-10, TNF-α and GAPDH amplification.
Shi B., Huang Y., Ni J., Chen J., Wei J., Gao H., Li L., Zhou Z., Wang Y., Xu Y., Xu Z., Mao J, & Fan G. (2020). Qi Dan Li Xin pill improves chronic heart failure by regulating mTOR/p70S6k-mediated autophagy and inhibiting apoptosis. Scientific Reports, 10, 6105.
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9

Liver Gene Expression Analysis

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The CFX96 real-time PCR detection system was used for real-time quantitative PCR (Bio-Rad, Munich, Germany). TRIZOL reagent (Takara Biotechnology Co., Ltd., Dalian, China) was used to extract total RNA from liver tissue, and Superscript II reverse transcriptase was used to prepare first strand cDNA (Roche, Basel, Switzerland). SYBR Green I PCR Master Mix (Vazyme Biotech Co.,Ltd, Nanjing, China) was used to determine mRNA levels, which were then calculated using the 2−ΔΔCT method. Sangon Biotech (Shanghai, China) designed specific primers (Table S1) for inflammatory genes (TNF-α, IL-1β, IL-6, and IL-10), antioxidant genes (SOD-1, SOD-2), and apoptosis genes (Bcl-2, Bax, and Caspase-3) based on the NCBI database sequence (Table S1).
Sui Y., Lu Y., Zuo S., Wang H., Bian X., Chen G., Huang S., Dai H., Liu F, & Dong H. (2022). Aflatoxin B1 Exposure in Sheep: Insights into Hepatotoxicity Based on Oxidative Stress, Inflammatory Injury, Apoptosis, and Gut Microbiota Analysis. Toxins, 14(12), 840.
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10

Retrovirus-specific Microarray Analysis

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Reverse transcription of mRNA was performed with 1μg total RNA using Superscript II (Roche Diagnostics, Mannheim) according to the manufacturers protocol. Three independent RNA isolations were used for microarray analysis. Amplification and labeling of the hybridization probes by MOP PCR, DNA microarray preparation, hybridization and post-processing of retrovirus-specific microarrays were performed as described previously [46 (link),53 (link)]. Exclusively arrays showing reproducible hybridization patterns in triplicate subarrays were further evaluated. Hybridized microarrays were scanned using an Affymetrix Scanner GMS 418 (laser power settings, 100%; gain, 50%), and false color mapping was used for image visualization.
Vincendeau M., Göttesdorfer I., Schreml J.M., Wetie A.G., Mayer J., Greenwood A.D., Helfer M., Kramer S., Seifarth W., Hadian K., Brack-Werner R, & Leib-Mösch C. (2015). Modulation of human endogenous retrovirus (HERV) transcription during persistent and de novo HIV-1 infection. Retrovirology, 12, 27.
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