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Rnaeasy rna purification kit

Manufactured by Qiagen
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The RNAeasy RNA purification kit is a product designed for the extraction and purification of ribonucleic acid (RNA) from various biological samples. It utilizes a silica-based membrane technology to efficiently capture and elute high-quality RNA.

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Lab products found in correlation

Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (3 mentions) Power sybr green rna to ct 1 step kit, by Thermo Fisher Scientific (2 mentions) Nanodrop spectrophotometer, by Thermo Fisher Scientific (2 mentions) Abi prism sds software, by Thermo Fisher Scientific (1 mentions) Bioanalyzer 2100 system, by Agilent Technologies (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Pcr master mix, by Thermo Fisher Scientific (1 mentions) Superscript 3, by Thermo Fisher Scientific (1 mentions) 7900ht fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Maxima probe rox qpcr master mix, by Thermo Fisher Scientific (1 mentions) Abi prism 7900, by Thermo Fisher Scientific (1 mentions)

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RNAeasy purification kit RNA purification kit RNAeasy kit RNAeasy

6 protocols using rnaeasy rna purification kit

1

Quantitative RT-PCR Analysis of Colon Tissue

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RNA was extracted from snap-frozen colon tissue using the RNAeasy RNA Purification Kit (Qiagen) including on-column DNase digestion. qPCR was performed using a Power SYBR Green RNA-to-CT 1-Step Kit (Applied Biosystems) according to the manufacturer’s instructions. Reactions were performed in 25 μl containing 50 ng RNA, 12.5 μl SYBR Green mix, 0.16 μl RT mix, and 0.2 μM primer (for sequences see Supplementary Table 4). Program: 30 min at 48 °C; 10 min at 95 °C; followed by 40 cycles of 15 s at 95 °C/60 s at 60 °C. For each oligonucleotide pair and RNA sample, the reaction was performed in triplicate. The amplification plots obtained from the RT-PCR were analyzed with StepOne™ Real-Time PCR Software v2.2. The expression levels of the target genes were normalized to the levels of glyceraldehyde-3-phosphate dehydrogenase gene expression in each individual sample.
Harnack C., Berger H., Antanaviciute A., Vidal R., Sauer S., Simmons A., Meyer T.F, & Sigal M. (2019). R-spondin 3 promotes stem cell recovery and epithelial regeneration in the colon. Nature Communications, 10, 4368.
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2

Quantitative RT-PCR Analysis of Gene Expression

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RNA was extracted from snap-frozen corpus tissue using the RNAeasy RNA Purification Kit (QIAGEN) including on-column DNase digestion. qPCR was performed using a Power SYBR Green RNA-to-CT 1-Step Kit (Applied Biosystems) according to the manufacturer’s instructions. Reactions were performed in a reaction mix of 25 μl containing 50 ng RNA, 12.5 μl SYBR Green mix, 0.16 μl RT mix, and 0.2 μM primer. Reactions were performed for 30 minutes at 48°C followed by 10 minutes at 95°C and then 40 cycles of 15 seconds at 95°C/60 seconds at 60°C.
For primers used in this study, see Table 1.
For each oligonucleotide pair and RNA sample, the reaction was performed in duplicate. The amplification plots obtained from RT-PCR were analyzed with the ABI Prism SDS Software package, version 2.2.2 (Applied Biosystems). The expression levels of the target genes were normalized to the levels of Gapdh expression in each sample.
Fischer A.S., Müllerke S., Arnold A., Heuberger J., Berger H., Lin M., Mollenkopf H.J., Wizenty J., Horst D., Tacke F, & Sigal M. (2022). R-spondin/YAP axis promotes gastric oxyntic gland regeneration and Helicobacter pylori–associated metaplasia in mice. The Journal of Clinical Investigation, 132(21), e151363.
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3

RNA Extraction and Purification Protocol

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At each designated time point of the experiment, the parasites were washed off the PHCM cells and the cells were lysed in RNA extraction lysis buffer as described by the manufacturer. Total RNA was purified with the RNAeasy RNA purification kit (Qiagen, Valencia, CA, USA). The RNA purification step included an on column DNase treatment step to eliminate traces of genomic DNA as described by the manufacturer (Qiagen). The quality of the purified RNA was analyzed using the Bioanalyzer 2100 system (Agilent Technologies, Santa Clara, CA, USA). The quality of purified RNA was assessed by RNA Integrity Number (RIN) and samples with a RIN of at least 7 were used for further analysis.
Udoko A.N., Johnson C.A., Dykan A., Rachakonda G., Villalta F., Mandape S.N., Lima M.F., Pratap S, & Nde P.N. (2016). Early Regulation of Profibrotic Genes in Primary Human Cardiac Myocytes by Trypanosoma cruzi. PLoS Neglected Tropical Diseases, 10(1), e0003747.
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4

RNA Extraction and qRT-PCR Analysis

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Total RNA was extracted from islets and INS-1 832/13 cells using RNAeasy RNA purification kit (Qiagen GmbH, Hilden, Germany). Equal quantities of total RNA were reverse transcribed using RevertAid™ First-Strand cDNA synthesis kit (Fermentas, Vilnius, Lithuania). qRT-PCR was performed on ViiA7 qRT-PCR system, using predesigned TaqMan Gene Expression assays (PE Applied Biosystems, Foster City, CA, USA). Gene expression was determined by the absolute quantification method and normalized to the expression of Hprt.
Nicholas L.M., Valtat B., Medina A., Andersson L., Abels M., Mollet I.G., Jain D., Eliasson L., Wierup N., Fex M, & Mulder H. (2017). Mitochondrial transcription factor B2 is essential for mitochondrial and cellular function in pancreatic β-cells. Molecular Metabolism, 6(7), 651-663.
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5

Quantitative Real-Time PCR for Gene Expression

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Total RNA was extracted from αTC1-6 and INS-1 832/13 cells using RNAeasy RNA purification kit (Qiagen GmbH, Hilden, Germany). RNA concentrations were determined using a NanoDrop Spectrophotometer (Thermo Scientific). Equal quantities of total RNA were reverse transcribed using RevertAid™First-Strand cDNA synthesis kit (Fermentas, Vilnius, Lithuania) or Superscript III (Life Technologies) in reactions containing 500 ng or 1000 ng of total RNA. Quantitative real-time PCR (Q-PCR) was performed with 7900 HT Fast Real-Time PCR system (Applied Biosystems). The PCR reaction mix consisted of first-strand cDNA template, primer pairs (for details, see supplementary table 1), 6-carboxyfluorescein/quencher probes (Bioresearch Technologies, CA and Applied Biosystems), and PCR Master mix (Applied Biosystems) and was carried out as previously described [40 (link)]. Expression of selected targets was compared with that of hypoxanthine-guanine phosphoribosyl transferase (Hprt) measured on the same sample in parallel on the same plate, using the comparative Ct method.
Stamenkovic J.A., Andersson L.E., Adriaenssens A.E., Bagge A., Sharoyko V.V., Gribble F., Reimann F., Wollheim C.B., Mulder H, & Spégel P. (2015). Inhibition of the malate-aspartate shuttle in mouse pancreatic islets abolishes glucagon secretion without affecting insulin secretion. The Biochemical journal, 468(1), 49-63.
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6

Gene Expression in Diabetic Rat Islets

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Isolated islets from DRLyp/Lyp (n = 6, 3M/3F) and control (n = 7, 3M/4F) rats were frozen (−80°C) after isolation or after 5–7 days in culture (37°C, 5% CO2 in RPMI medium, 5.6 mmol/l glucose, 10% FBS + penicillin [100 units/ml]–streptomycin [100 μg/ml]). Total RNA was extracted (RNAeasy RNA purification kit; Qiagen, Hilden, Germany) and equal quantities of RNA were reverse transcribed (RevertAid First-Strand cDNA synthesis kit; Fermentas, Vilnius, Lithuania). mRNA levels were quantified (Maxima Probe/ROX qPCR Master Mix; Fermentas, Thermo Scientific, Helsingborg, Sweden) using an ABI PRISM 7900 (Applied Biosystems ViiA Real Time PCR System; Life Technologies, Foster City, CA, USA), using probes for Il1b (ID no. Rn00580432), Tnf-α (also known as Tnf) (ID no. Rn01525859), Ifng (ID no. Rn00594078) and Glut2 (also known as Slc2a2) (ID no. Rn00563565) (Applied Biosystems). Samples were run in triplicate and the transcript quantity was normalised to the geometric mean of mRNA levels of the reference genes (Applied Biosystems) Ppia (ID no. Rn00690933), Polr2a (ID no. Rn01752026) and Hprt (also known as Hprt1) (ID no. Rn01527840), using the formula 2(minCt – sampleCt).
Medina A., Parween S., Ullsten S., Vishnu N., Siu Y.T., Quach M., Bennet H., Balhuizen A., Åkesson L., Wierup N., Carlsson P.O., Ahlgren U., Lernmark Å, & Fex M. (2017). Early deficits in insulin secretion, beta cell mass and islet blood perfusion precede onset of autoimmune type 1 diabetes in BioBreeding rats. Diabetologia, 61(4), 896-905.
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