Log In Sign up
The largest database of trusted experimental protocols

Rneasy plus kit

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The RNeasy Plus Kit is a laboratory equipment designed for the purification of total RNA from a variety of biological samples. It utilizes a silica-membrane-based technology to efficiently capture and purify RNA molecules.

Automatically generated - may contain errors

Lab products found in correlation

Rneasy plus kit, by Qiagen (2 mentions) Brilliant 3 ultra fast qpcr master mix, by Agilent Technologies (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Ns500, by Illumina (2 mentions) Mxaria, by Agilent Technologies (2 mentions) Superscript 3 first strand synthesis system, by Thermo Fisher Scientific (2 mentions) Taqman probe, by Thermo Fisher Scientific (2 mentions) Superscript 3, by Thermo Fisher Scientific (2 mentions) One step plus system, by Thermo Fisher Scientific (2 mentions) Iscript cdna synthesis kit, by Bio-Rad (2 mentions) Cfx384 real time system, by Bio-Rad (1 mentions) Quantitect sybr green pcr kit, by Qiagen (1 mentions) 7300 real time pcr system, by Thermo Fisher Scientific (1 mentions) Pcr 2720 thermal cycler, by Thermo Fisher Scientific (1 mentions) Sybr green, by Bio-Rad (1 mentions) Cfx96 real time system, by Bio-Rad (1 mentions) Ssofast evagreen supermix, by Bio-Rad (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Purelink rna columns, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

RNeasy kit (131 citations) RNeasy Plus Mini Kit (46 citations) RNeasy plus micro kit (19 citations) RNeasy Plus Universal Tissue Mini Kit (6 citations) PLUSTM (3 citations) RNeasy Mini Plus extraction kit (3 citations) RNeasy Plus Isolation Kit (2 citations)

10 protocols using rneasy plus kit

1

RNA-seq Analysis of Gene Expression

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was extracted from cell pellets using the RNeasy Plus kit (QIAGEN). RNA was checked for quality using TapeStation and Qubit before library preparation using Kappa stranded mRNA Hyper Prep and sequencing using Illumina NS500. Reads were mapped to Hg19 using STAR alignment (Dobin et al., 2013 (link)) and counts were generated using HTseq (Anders et al., 2015 (link)). Differential gene expression analysis was perform using DESeq2 (Love et al., 2014 (link)) and heatmaps were generated using R package pheatmap. Mean counts were generated using DESeq2 mean-ratio normalization method. Gene set enrichment analysis was conducted using GSEA software (https://www.broad.mit.edu/gsea/downloads.jsp) (Mootha et al., 2003 (link); Subramanian et al., 2005 (link)). RNA-seq datasets have been deposited in the GEO database (accession number GSE154545). Gene annotations in Figure 5D for G1/S and G2/M genes are from Schade et al. (2019) (link). RT-qPCR to validate RNA expression data was done using cDNA generated from total RNA isolated by the RNeasy Plus kit and the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qPCR was performed on the cDNA using the primers listed in Table S3 and Brilliant III Ultra-Fast qPCR Master Mix (Agilent) using a MxAria (Agilent)
Branigan T.B., Kozono D., Schade A.E., Deraska P., Rivas H.G., Sambel L., Reavis H.D., Shapiro G.I., D’Andrea A.D, & DeCaprio J.A. (2021). MMB-FOXM1-driven premature mitosis is required for CHK1 inhibitor sensitivity. Cell reports, 34(9), 108808.
+ Open protocol
+ Expand
2

RNA-seq Analysis of Gene Expression

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was extracted from cell pellets using the RNeasy Plus kit (QIAGEN). RNA was checked for quality using TapeStation and Qubit before library preparation using Kappa stranded mRNA Hyper Prep and sequencing using Illumina NS500. Reads were mapped to Hg19 using STAR alignment (Dobin et al., 2013 (link)) and counts were generated using HTseq (Anders et al., 2015 (link)). Differential gene expression analysis was perform using DESeq2 (Love et al., 2014 (link)) and heatmaps were generated using R package pheatmap. Mean counts were generated using DESeq2 mean-ratio normalization method. Gene set enrichment analysis was conducted using GSEA software (https://www.broad.mit.edu/gsea/downloads.jsp) (Mootha et al., 2003 (link); Subramanian et al., 2005 (link)). RNA-seq datasets have been deposited in the GEO database (accession number GSE154545). Gene annotations in Figure 5D for G1/S and G2/M genes are from Schade et al. (2019) (link). RT-qPCR to validate RNA expression data was done using cDNA generated from total RNA isolated by the RNeasy Plus kit and the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qPCR was performed on the cDNA using the primers listed in Table S3 and Brilliant III Ultra-Fast qPCR Master Mix (Agilent) using a MxAria (Agilent)
Branigan T.B., Kozono D., Schade A.E., Deraska P., Rivas H.G., Sambel L., Reavis H.D., Shapiro G.I., D’Andrea A.D, & DeCaprio J.A. (2021). MMB-FOXM1-driven premature mitosis is required for CHK1 inhibitor sensitivity. Cell reports, 34(9), 108808.
+ Open protocol
+ Expand
3

Quantitative Analysis of Osteogenic Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
RNA was isolated with an RNeasy Plus Kit and reverse transcribed into cDNA using the SuperScript III First-Strand Synthesis System (Invitrogen). Gene expression was measured using qRT-PCR. Reactions included 25 ng cDNA per 10μl with QuantiTect SYBR Green PCR Kit (Qiagen, Valencia, CA) and the CFX384 Real-Time System (BioRad, Hercules, CA). Transcript levels were normalized to the housekeeping gene Gapdh. Gene expression levels were quantified using the 2ΔΔCt method. Gene specific primer sequences: Gapdh (Forward: 5′-GGGAAG CCCATCACCATCTT, Reverse: 5′-GCCTCACC CCATTTG ATGTT), Osteocalcin (Bglap) (Forward: 5′-CCTGAGTCTGACAAAGCCTTCA, Reverse: 5′-GCCGGAGTCTGTTCACTACCTT), and Alkaline phosphatase (Alpl) (Forward: 5′-CACAGATTCCCAAAGCACCT, Reverse: 5′-GGGATGGAGGAGAGAAGGTC).
Kremer K.N., Dudakovic A., McGee-Lawrence M.E., Philips R.L., Hess A.D., Smith B.D., van Wijnen A.J., Karp J.E., Kaufmann S.H., Westendorf J.J, & Hedin K.E. (2014). Osteoblasts Protect AML Cells from SDF-1-Induced Apoptosis. Journal of cellular biochemistry, 115(6), 1128-1137.
+ Open protocol
+ Expand
4

Gene Expression Analysis via qPCR

Check if the same lab product or an alternative is used in the 5 most similar protocols
mRNA was isolated (Qiagen RNeasy Plus Kit) and transcribed to cDNA (Invitrogen SuperScript III). Gene expression was analyzed using Taqman probes and the OneStep Plus system (Applied Biosystems). Each biological sample was analyzed in experimental replicate (n=2 repeated wells of the qPCR reaction) and the Ct value of each replicate was averaged and handed as n=1 unique biologic sample. Expression for each sample was normalized to β-Actin (ACTA1) gene expression (ΔCt) and relative to peripheral lung tissue control samples (ΔΔCt), and fold change calculated by 2-ΔΔCt (24 (link)). A total of n=3 unique biological samples were analyzed for each experiment.
Gilpin S.E., Li Q., Evangelista-Leite D., Ren X., Reinhardt D.P., Frey B.L, & Ott H.C. (2017). Fibrillin-2 and Tenascin-C Bridge the Age Gap in Lung Epithelial Regeneration. Biomaterials, 140, 212-219.
+ Open protocol
+ Expand
5

Quantitative PCR for RNAi Knockdown Efficiency

Check if the same lab product or an alternative is used in the 5 most similar protocols
Quantitative PCR was used to assay efficiency of RNAi knockdowns according to standard techniques. Briefly, total RNA was isolated from cells using a Qiagen RNeasy Plus kit and then converted to cDNA using the Invitrogen SuperScript III First-Strand Synthesis System for RT-PCR. Primers for qPCR were designed using the Primer-BLAST website (http://www.ncbi.nlm.nih.gov/tools/primer-blast/). Reactions were set up using KAPA SYBR FAST qPCR kits and run on an Applied Biosystems 7300 Real-Time PCR System.
Senaratne T.N., Joyce E.F., Nguyen S.C, & Wu C.T. (2016). Investigating the Interplay between Sister Chromatid Cohesion and Homolog Pairing in Drosophila Nuclei. PLoS Genetics, 12(8), e1006169.
+ Open protocol
+ Expand
6

Quantitative Gene Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
We conducted RT-PCR to verify gene-level changes in the CON, DSS, and H-BBr groups. Extracted total RNA from colon tissues using RNeasy Plus Kit (Invitrogen, New York, United States). Following the manufacturer’s instructions, reverse transcription performing by using the BioRT Master HisSensi cDNA First-Strand Synthesis kit (Shanghai Roche, China). PCR2720 thermal cycler (Applied Biosystems, United States) and ABI7500 PCR system perform RT-qPCR. The fluorescence reagent used in this experiment was SYBR Green I, and the cycling conditions refer to Supplementary Figure 2. The primers used for PCR amplification which listed in Supplementary Table 2.
Cao J., Chen M., Xu R, & Guo M. (2022). Therapeutic Mechanisms of Berberine to Improve the Intestinal Barrier Function via Modulating Gut Microbiota, TLR4/NF-κ B/MTORC Pathway and Autophagy in Cats. Frontiers in Microbiology, 13, 961885.
+ Open protocol
+ Expand
7

Quantitative Real-Time PCR Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was isolated using the RNeasy plus kit (Invitrogen) according to the manufacturer’s instruction. cDNA was synthesized from total RNA using the ProtoScript First Strand cDNA Synthesis Kit (NEBioLabs). Quantitative real-time PCR was performed using SYBR Green (BioRad). The fold induction of each target gene was calculated using the comparative CT method (2-ΔΔCt). The Ct of each gene of interest was first normalized to the housekeeping gene GAPDH (ΔCt), and then to the control of the experiment (vehicle or DMSO treatment) to calculate the ΔΔCt., The oligonucleotide sequences used were the following:
Bolamperti S., Saito H., Heerdmann S., Hesse E, & Taipaleenmäki H. (2024). Tgif1-deficiency impairs cytoskeletal architecture in osteoblasts by activating PAK3 signaling. eLife, 13, RP94265.
+ Open protocol
+ Expand
8

Quantifying Gene Expression in Lung Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
mRNA was isolated (Qiagen RNeasy Plus Kit) and transcribed to cDNA (Invitrogen SuperScript III). Gene expression was analyzed using Taqman probes and the OneStep Plus system (Applied Biosystems). Each biological sample was analyzed in experimental replicate (n=2 repeated wells of the qPCR reaction) with the Ct value of each replicate averaged and handed as n=1 unique biologic sample. Expression for each sample was normalized to β-Actin (ACTA1) gene expression (ΔCt) and relative to cadaveric peripheral lung tissue control samples (ΔΔCt), with fold change calculated by 2−ΔΔCt (33 (link)).
Gilpin S.E., Charest J.M., Ren X., Tapias L.F., Wu T., Da Silva D.E., Mathisen D.J, & Ott H.C. (2016). Regenerative Potential of Human Airway Stem Cells in Lung Epithelial Engineering. Biomaterials, 108, 111-119.
+ Open protocol
+ Expand
9

Quantitative Gene Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
For cultured haSMCs or haECs, RNA was isolated with the RNeasy Plus Kit (Life Technologies). For newborn mice, PBS was perfused through the left ventricle, the entire aorta from the root to the iliac arteries was dissected, and aortic RNA was extracted with mechanical homogenization in TRIzol (Invitrogen) and PureLink RNA columns (Invitrogen). The isolated RNA was reverse transcribed with the iScript cDNA Synthesis Kit (Bio-Rad), and qRT-PCR was performed on a CFX96 Real-Time System (Bio-Rad) using SsoFast EvaGreen supermix (Bio-Rad) and primer pairs as per Supplemental Table 3. Normalized mRNA levels are relative to 18S rRNA for cultured cells and to 18S rRNA or Gapdh for murine aortas.
Dave J.M., Chakraborty R., Ntokou A., Saito J., Saddouk F.Z., Feng Z., Misra A., Tellides G., Riemer R.K., Urban Z., Kinnear C., Ellis J., Mital S., Mecham R., Martin K.A, & Greif D.M. (2022). JAGGED1/NOTCH3 activation promotes aortic hypermuscularization and stenosis in elastin deficiency. The Journal of Clinical Investigation, 132(5), e142338.
+ Open protocol
+ Expand
10

Vascular Gene Expression Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Descending aortas were dissected from mouse embryos and snap frozen in liquid nitrogen. The RNA of individual aortas of specific Eln genotypes was isolated with the RNeasy Plus kit (Life Technologies), and this RNA (0.5 µg) was reverse transcribed with the iScript cDNA Synthesis kit (Bio-Rad Laboratories). Expression levels of selected genes were determined by quantitative PCR and normalized to Gapdh. The following forward and reverse primer pairs were used for these genes (encoded protein): Acta2 (SMA), 5′-GTCCCAGACATCAGGGAGTAA-3′ and 5′-TCGGATACTTCAGCGTCAGGA-3′; Tagln (SM22-α), 5′-CAACAAGGGTCCATCCTACGG-3′ and 5′-ATCTGGGCGGCCTACATCA-3′; Myh11 (SMMHC), 5′-AAGCTGCGGCTAGAGGTCA-3′ and 5′-CCCTCCCTTTGATGGCTGAG-3′; Itgb1 (Integrin β1), 5′-CTCCAGAAGGTGGCTTTGATGC-3′ and 5′-GTGAAACCCAGCATCCGTGGAA-3′; Itgb3 (Integrin β3), 5′-GTGAGTGCGATGACTTCTCCTG-3′ and 5′-CAGGTGTCAGTGCGTGTAGTAC-3′; and Gapdh (GAPDH), 5′-CATCACTGCCACCCAGAAGACTG-3′ and 5′-ATGCCAGTGAGCTTCCCGTTCAG-3′.
Misra A., Sheikh A.Q., Kumar A., Luo J., Zhang J., Hinton R.B., Smoot L., Kaplan P., Urban Z., Qyang Y., Tellides G, & Greif D.M. (2016). Integrin β3 inhibition is a therapeutic strategy for supravalvular aortic stenosis. The Journal of Experimental Medicine, 213(3), 451-463.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!