Quantitect cdna reverse transcription kit

Manufactured by Qiagen

Sourced in Germany

The QuantiTect cDNA Reverse Transcription Kit is a laboratory product designed for the efficient conversion of RNA into complementary DNA (cDNA). The kit includes reagents and enzymes necessary for this reverse transcription process, enabling the generation of high-quality cDNA from various RNA samples.

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Lab products found in correlation

Nanodrop 1000, by Thermo Fisher Scientific (4 mentions) Taqman probe, by Thermo Fisher Scientific (3 mentions) Rneasy plus kit, by Qiagen (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions) Cfx maestro software, by Bio-Rad (1 mentions) Mm99999915 g1, by Thermo Fisher Scientific (1 mentions) Mm03024075 m1, by Thermo Fisher Scientific (1 mentions) Taqman chemistry, by Thermo Fisher Scientific (1 mentions) 7500 fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Rneasy micro kit, by Qiagen (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Qiaamp dna stool mini kit, by Qiagen (1 mentions) Sybr green pcr master mix, by Qiagen (1 mentions) High pure rna tissue kit, by Roche (1 mentions) Taqman gene expression real time qpcr assays, by Thermo Fisher Scientific (1 mentions) C100 thermal cycler, by Bio-Rad (1 mentions)

Spelling variants of this product

QuantiTect (227 citations) Reverse transcription kit (213 citations) QuantiTect kit (107 citations) QuantiTect Reverse Transcription (82 citations) QuantiTect Transcription Kit (12 citations) QuantiTect Reverse Transcription cDNA synthesis kit (3 citations) Quantitect Reverse Transcription Kit for cDNA synthesis for PCR (3 citations) QuantiTect reverse kit (2 citations) QuantiTect Reverse Transcription Kit cDNA synthesis kit (2 citations)

5 protocols using quantitect cdna reverse transcription kit

Quantitative RT-PCR for Gene Expression

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Total RNA was extracted using RNeasy (RNeasy plus kit, Qiagen, Milano, Italy), quantified using Nanodrop 1000 (Thermo Scientific) and 500 ng were reverse transcribed, prior to gDNA removal, to obtained cDNA using the QuantiTect cDNA reverse transcription kit (Qiagen). Quantitative real-time polymerase chain reaction (qRT-PCR) was performed using the CFX96 Touch Real-Time PCR Detection System (Biorad) using Taqman gene expression assay (Applied Biosystem, Thermo Fisher Scientific). The probes used are HKII (Hs0060686_m1) and GLUT1 (Hs00892681_m1). Data were normalized on Glyceraldehyde 3-Phosphate Dehydrogenase GAPDH (Hs02786624_g1) and TATA-Box Binding Protein TBP (Hs00427620_m1). The relative quantity was determined using ΔΔCt by the CFX Maestro software (BioRad).

Lorito N., Bacci M., Smiriglia A., Mannelli M., Parri M., Comito G., Ippolito L., Giannoni E., Bonechi M., Benelli M., Migliaccio I., Malorni L., Chiarugi P, & Morandi A. (2020). Glucose Metabolic Reprogramming of ER+ Breast Cancer in Acquired Resistance to the CDK4/6 Inhibitor Palbociclib. Cells, 9(3), 668.

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RNA Expression Analysis of m6A Regulators

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For qRT PCR, 1 μg of RNA was retrotranscribed using the Quantitect cDNA reverse transcription kit (Qiagen). Quantitative real-time PCR was performed using PrimeTime® Gene Expression Master Mix and the following probes (all IDT): Mettl3 (Mm.PT.58.10309074), Mettl14 (Mm.PT.58.33500780), Mettl16 (Mm.PT.58.9941116), Alkbh5 (Mm.PT.58.6928987), Fto (Mm.PT.58.32888407), and Pcif1 (Mm.PT.58.9572065). TaqMan probes for housekeeping genes Gapdh (Mm99999915_g1) and Hprt (Mm03024075_m1), were from Thermo Fisher.

Jabs S., Biton A., Bécavin C., Nahori M.A., Ghozlane A., Pagliuso A., Spanò G., Guérineau V., Touboul D., Giai Gianetto Q., Chaze T., Matondo M., Dillies M.A, & Cossart P. (2020). Impact of the gut microbiota on the m6A epitranscriptome of mouse cecum and liver. Nature Communications, 11, 1344.

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Circadian Rhythms Gene Expression Analysis

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The tissue was processed for RNA extraction using Qiazol extraction and RNeasy Micro Kit (QIAGEN). Concentrations of mRNA were determined using spectrophotometry with samples diluted to the same final concentration. Immediately following extraction, mRNA was treated with Ambion DNase treatment and removal (Life Technologies) and cDNA synthesis was performed with QuantiTect^® cDNA reverse transcription kit (QIAGEN). Reverse-transcription qPCR assays were performed using TaqMan chemistry commercially available validated assays from Life Technologies. Transcripts were amplified on a 7500 Fast Real-Time PCR System by Applied Biosystems (Life Technologies) using TaqMan probes for Rn18s (control gene; Mm03928990_g1), Per1 (Mm00501813_m1), Per2 (Mm00478099_m1), Bmal1 (Arntl1) (Mm00500226_m1), Nr1d1 (Rev-erbα) (Mm00520708_m1), Clock (Mm00455950_m1), Cckar (Mm00438060_m1), Cckbr (Mm00432329_m1). Samples were run in triplicate with a standardized 5 ng (2.5 ng / μL × 2 μL) of cDNA and compared using the ^ΔΔCT method of relative quantification, with Rn18s used as the control housekeeping gene and ZT0 values as the relative target. Kruskal-Wallis test with post-hoc multiple comparison was performed for statistical analysis.

Barthelmebs L., Divies C, & Cavin J.F. (2000). Knockout of the p-Coumarate Decarboxylase Gene from Lactobacillus plantarum Reveals the Existence of Two Other Inducible Enzymatic Activities Involved in Phenolic Acid Metabolism. Applied and Environmental Microbiology, 66(8), 3368-3375.

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Antimicrobial Effects of Herbal Extract

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Herbal medicine markets in Mashhad, Iran, provided fresh aerial parts of A. millefolium. Sigma Aldrich (Germany) supplied soybean lecithin with a purity of 99%. As a food-borne pathogen, C. jejuni (RTCC 1097) was provided from Razi Vaccine and Serum Research Institute, Karaj, Iran (Gharib Naseri et al., 2012 ; Ebrahimi et al., 2016 (link)). As part of the gene expression analysis, the RNeasy Mini kit from Qiagen (Hilden, Germany), SYBR Green PCR Master Mix from Qiagen (Hilden, Germany), cDNA Quantitect Reverse Transcription Kit (Qiagen, Hilden, Germany), and SYBR Green PCR Master Mix from Qiagen (Hilden, Germany) were utilised. In this study, DNA was extracted using the QIAamp DNA Stool Mini Kit from Qiagen, Hilden, Germany. Besides these, Merck (Germany) provided the rest of the reagents.

Nateghi N., Karimi E, & Oskoueian E. (2022). Nanoliposome-Encapsulated and Non-Encapsulated Phenolics From Achillea millefolium and Their Biological Function in Mice Challenged by Campylobacter jejuni: A Comparative Study. Frontiers in Molecular Biosciences, 8, 832022.

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Quantitative PCR Analysis of Retinal Genes

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Total RNA from neuroretina and RPE samples was extracted using High Pure RNA tissue kit (Roche) according to the manufacturer’s instructions. After spectrophotometric quantification of RNA using NanoDrop1000 (Thermo Fisher Scientific), reverse transcription was carried out using cDNAQuantiTect® reverse transcription kit (Qiagen, Hilden, Germany), according to manufacturer’s instructions. cDNA amplification was performed by using 1 μg of RNA as template. Approximately 100 ng of cDNA was used for qPCR. Specific primers for Prpf31 (Mm01329809_m1, Thermo Fisher Scientific), Recoverin (Mm00501325_m1, Thermo Fisher Scientific), Rpe65 (Mm00504133_m1, Thermo Fisher Scientific) and Hspa4l (Mm00495441_m1, Thermo Fisher Scientific) were used. The qPCR was performed using TaqMan® Gene Expression Real Time qPCR assays (Life-Technologies, California, USA) according to the manufacturer’s instructions, using a Thermal Cycler C100 (Bio-Rad). The average cycle threshold (CT) of fluorescence units was used to analyze the mRNA levels. Prpf31, Recoverin, Rpe65 and Hspa4l mRNA levels were normalized by Gapdh RNA levels. Quantification was calculated as: mRNA levels (percent of control) = 2^Δ(CT) with Δ (CT) = CT_{(Prpf31/Recoverin/Rpe65/Hspa4l)}- CT_(Gapdh).

Valdés-Sánchez L., Calado S.M., de la Cerda B., Aramburu A., García-Delgado A.B., Massalini S., Montero-Sánchez A., Bhatia V., Rodríguez-Bocanegra E., Diez-Lloret A., Rodríguez-Martínez D., Chakarova C., Bhattacharya S.S, & Díaz-Corrales F.J. (2019). Retinal pigment epithelium degeneration caused by aggregation of PRPF31 and the role of HSP70 family of proteins. Molecular Medicine, 26, 1.

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