Total rna extraction kit

Manufactured by Merck Group

Sourced in United States

The Total RNA Extraction Kit is a comprehensive solution for the isolation and purification of high-quality total RNA from a variety of biological samples. The kit utilizes a proprietary silica-based membrane technology to efficiently capture and purify RNA molecules, including mRNA, rRNA, and small non-coding RNAs. The extracted RNA is ready for downstream applications such as reverse transcription, real-time PCR, northern blotting, and RNA sequencing.

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Lab products found in correlation

Reverse transcriptase, by Promega (3 mentions) Cdna synthesis kit, by Takara Bio (1 mentions) Tb green premix ex taq kit, by Takara Bio (1 mentions) Quantstudio 6 flex real time system, by Thermo Fisher Scientific (1 mentions) Reverse transcription kit, by Merck Group (1 mentions) Agarose gel, by Thermo Fisher Scientific (1 mentions) Rotor gene 6000, by Qiagen (1 mentions)

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RNA extraction kit (24 citations)

6 protocols using total rna extraction kit

RNA Extraction and Transcriptome Analysis

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A total RNA extraction kit (Axygen, Sigma‐Aldrich) was used to extract total RNA from the NPCs. The first strand complementary DNA (cDNA) synthesis kit (TAKARA, Beijing, China) was used to reverse transcribe the total RNA to first strand cDNA. Quantitative PCR was performed using the TB‐Green premix Ex Taq kit (TAKARA) system and a QuantStudio 6 Flex real‐time system (Thermo Fisher Scientific). The 2^{^(−ΔΔCt)} and 2^{^(−ΔCt)} methods were used to calculate relative gene expression. β‐actin and GAPDH were used as internal references. Primer sequences were designed using BLAST (NCBI, Bethesda, USA).
Extracted RNA was assayed via RNA (transcriptome) sequencing by Wuhan Huada Gene Technology Co., Ltd. (China) and analyzed for the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, Gene Ontology (GO) cellular components, and mRNA relative expression using fragments per kilobase million (FPKM) on the Mybgi platform (Wuhan Huada Gene Technology, https://mybgi.bgi.com/tech/login).

Zhou T., Yang X., Chen Z., Yang Y., Wang X., Cao X., Chen C., Han C., Tian H., Qin A., Fu J, & Zhao J. (2022). Prussian Blue Nanoparticles Stabilize SOD1 from Ubiquitination‐Proteasome Degradation to Rescue Intervertebral Disc Degeneration. Advanced Science, 9(10), 2105466.

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Yihe-Tang Total RNA Extraction

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Yihe-Tang was purchased from the Affiliated Hospital of Inner Mongolia Minzu University (approval number: Z15021057). The total RNA extraction kit, reverse transcription kit, and other drugs were purchased from Sigma-Aldrich (St. Louis, MO).

Sa R., Feng C., Bai H., Yin X., Song L., Hu X., Xu R., Li X., Dong W, & Yang J. (2022). Inhibitory Effects of Mongolian Medicine Yihe-Tang on Continuous Darkness Induced Liver Steatosis in Zebrafish. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 5794655.

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Quantifying Tomato Gene Expression

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Total RNA extraction kit (Sigma-Aldrich, Munich, Germany) is used for the isolation of RNA from tomato leaves. Then the SuperScript cDNA synthesis kit is used for cDNAs formation. 18s gene was used as a reference gene in the relative expression of gene expression; primers sequence of catalase and heat shock proteins are in Table 1. The reaction is made of 2 µL of forward primer, 2 µL of reverse primer, 10 µL of SYBR Green Master Mix, 2 µL of the template, and sterile water for a total volume of 20 µL. Livak equation 2^−ΔΔCt are used for the calculation of relative gene expression [110 (link)].

Elkelish A., Qari S.H., Mazrou Y.S., Abdelaal K.A., Hafez Y.M., Abu-Elsaoud A.M., Batiha G.E., El-Esawi M.A, & El Nahhas N. (2020). Exogenous Ascorbic Acid Induced Chilling Tolerance in Tomato Plants Through Modulating Metabolism, Osmolytes, Antioxidants, and Transcriptional Regulation of Catalase and Heat Shock Proteins. Plants, 9(4), 431.

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Artemisia RNA Extraction and RT-PCR

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Total mRNA was isolated from 250 mg fresh leaves of Artemisia plants using Total RNA extraction kit (Sigma-Aldrich) according to the manufacturer’s protocol. The purified RNA was quantitated spectrophotometrically and analyzed on 1% agarose gel (Thermo Fisher Scientific, Waltham, MA, USA). Reverse transcription reactions were performed using oligo dT primer. The 20 μL reaction mixture contained 2.5 μL 5× buffer, 2.5 μL MgCl₂, primer (10 pml/μL), 2.5 μL 2.5 mM dNTPs (QIAGEN, Hilden, Germany), 4 μL from oligo (dT), 0.2 μL (5 Unit/μL) reverse transcriptase (Promega, Madison, WI, USA), and 2.5 μL RNA. RT-PCR amplification was performed in a thermal cycler PCR (QIAGEN, Germantown, MD, USA), programmed at 42 °C for one h and 72 °C for 20 min.

, & Alhaithloul H.A. (2019). Impact of Combined Heat and Drought Stress on the Potential Growth Responses of the Desert Grass Artemisia sieberi alba: Relation to Biochemical and Molecular Adaptation. Plants, 8(10), 416.

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Wheat mRNA Extraction and Quantification

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Total mRNA was isolated from 0.5 g wheat plant leaves from all treatment groups using a total RNA extraction kit (Sigma-Aldrich, St. Louis, MO, USA) based on the manufacturer’s method. The purified RNA was measured spectrophotometrically, and RNA reverse transcription was performed. The reaction mixture consisted of 2.5 μL 5× buffer, 2.5 μL 2.5 mM dNTPs, 2.5 μL MgCl₂, 4 μL oligo (dT), oligo dT primer (10 pmL/μL), 2.5 μL RNA and 0.2 μL (5 unit/μL) reverse transcriptase (Promega, Gutenbergring 10, 69190 Walldorf, Germany). Primers for specific genes and housekeeping genes were used in real-time analysis with a Rotor-Gene 6000 (Qiagen, Hilden, Germany). The relative gene expression was measured by following the 2^−ΔΔCt methods of Livak and Schmittgen [50 (link)]. Specific gene accession numbers and the sequences of specific primers designed for qRT-PCR are listed in Table S2.

Hasan M.M., Alharbi B.M., Alhaithloul H.A., Abdulmajeed A.M., Alghanem S.M., Al-Mushhin A.A., Jahan M.S., Corpas F.J., Fang X.W, & Soliman M.H. (2021). Spermine-Mediated Tolerance to Selenium Toxicity in Wheat (Triticum aestivum L.) Depends on Endogenous Nitric Oxide Synthesis. Antioxidants, 10(11), 1835.

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Leaf Transcriptome Analysis via RNA Extraction and RT-PCR

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According to the manufacturer’s protocol, a total RNA extraction kit (Sigma-Aldrich, St. Louis, MO, USA) was used for total mRNA extraction that was isolated from 0.1 g of leaf tissue. The isolated RNA was spectrophotometrically quantified and examined on a 1% agarose gel. RNA reverse transcription was carried out by following our previous method (Alhaithloul, 2019 (link)). The reaction mixture were included 10 as oligodT primer (10 pml/µL), 2.5 µL 5X buffer, 2.5 µL MgCl₂, 2.5 µL 2.5 mMdNTPs, 4 µl from oligo (dT), 0.2 µL (5 unit µL⁻¹) reverse transcriptase (Promega, Madison, WI, USA) and 2.5 µL RNA. The RT-PCR amplification was carried out in a thermal cycler PCR set to 42 °C for 1 h and 72 °C for 20 min.

Sakit ALHaithloul H.A., Khan M.I., Musa A., Ghoneim M.M., Aysh ALrashidi A., Khan I., Azab E., Gobouri A.A., Sofy M.R., El-Sherbiny M, & Soliman M.H. (2022). Phytotoxic effects of Acacia saligna dry leachates on germination, seedling growth, photosynthetic performance, and gene expression of economically important crops. PeerJ, 10, e13623.

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