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Rneasy micro extraction kit

Manufactured by Qiagen
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Sourced in Germany, France

The RNeasy Micro extraction kit is a laboratory equipment designed for the isolation and purification of high-quality total RNA from small amounts of starting material, such as cells or tissues. The kit utilizes a silica-membrane technology to efficiently capture and purify RNA, allowing for its use in various downstream applications.

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Lab products found in correlation

Abi 7500, by Thermo Fisher Scientific (4 mentions) Iscript cdna synthesis kit, by Bio-Rad (4 mentions) Quantstudio 6 flex real time pcr system, by Thermo Fisher Scientific (4 mentions) Ssoadvanced universal sybr green supermix, by Bio-Rad (3 mentions) Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (3 mentions) Superscript 2, by Thermo Fisher Scientific (2 mentions) Steponeplus, by Thermo Fisher Scientific (2 mentions) Cfx real time pcr machine, by Bio-Rad (2 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Kapa sybr fast abi prism 2 qpcr master mix, by Roche (2 mentions) Stepone real time pcr system, by Thermo Fisher Scientific (2 mentions) Quantitect reverse transcription kit, by Qiagen (2 mentions) High capacity rna to cdna kit, by Thermo Fisher Scientific (2 mentions) Stepone real time pcr systems software v2, by Thermo Fisher Scientific (2 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) Taqman universal master mix, by Thermo Fisher Scientific (2 mentions) Kapa sybr fast abi prism 2x qpcr master mix, by Roche (2 mentions) Nanodrop 2000 machine, by Thermo Fisher Scientific (2 mentions) Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

RNeasy kit (11240 citations) RNeasy Micro Kit (4852 citations) RNeasy extraction kit (334 citations) RNeasy Micro (65 citations) RNeasy Plus Micro extraction kit (10 citations) RNeasy Micro RNA extraction kit (9 citations) RNA Extraction RNeasy Plus Mini- or Micro-kit (2 citations)

27 protocols using rneasy micro extraction kit

1

RNA Extraction and qPCR Analysis

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RNA from cells or from lymph-nodes and footpads tissue was extracted using respectively a microRNeasy extraction kit (Qiagen) or TRIzol Reagent (Thermo Fisher Scientific), according to the manufacturer’s protocols. RNA (2 μg) was reverse transcribed using (200 U) Moloney murine leukemia virus reverse transcriptase (SuperScript II, Invitrogen). Subsequent real time PCR was performed on Step One Plus (Applied Biosystems) using Taq polymerase (Taq-Man Universal PCR master mix, Applied biosystems). All PCR data values were normalized to the expression of the hypoxanthine phosphoribosyltransferase (HPRT) gene.
Rasid O., Mériaux V., Khan E.M., Borde C., Ciulean I.S., Fitting C., Manoury B., Cavaillon J.M, & Doyen N. (2016). Cathepsin B-Deficient Mice Resolve Leishmania major Inflammation Faster in a T Cell-Dependent Manner. PLoS Neglected Tropical Diseases, 10(5), e0004716.
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2

RNA Extraction and qRT-PCR Analysis

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RNA was extracted from BMDCs or Gen2.2 using a microRNeasy extraction kit (Qiagen). A trace of genomic DNA was removed by RNAse free-DNAse set. RNA (2 µg) was reverse transcribed using (200 U) Moloney murine leukemia virus reverse transcriptase (SuperScript II, Invitrogen). Subsequent real time PCR was performed on Step One Plus (Applied Biosystems) using Taq polymerase or SYBER green (Taq-Man Universal or SYBER Green PCR master mix, Applied biosystems).
Khan M.E., Borde C., Rocha E.P., Mériaux V., Maréchal V., Escoll P., Goyard S., Cavaillon J.M., Manoury B, & Doyen N. (2014). TLR9 Activation Is Triggered by the Excess of Stimulatory versus Inhibitory Motifs Present in Trypanosomatidae DNA. PLoS Neglected Tropical Diseases, 8(11), e3308.
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3

Quantifying Extracellular Matrix Remodeling

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Cells were cultured on curved collagen gels for 48 hours; gels were then cut into three sections—one curved region and two plateau regions—using a razor blade (Sup. Fig. 1C). Curved and plateau regions were separated and then pooled across six collagen gels. The resulting curved or plateau samples were placed into 350 μL Buffer RLT (Qiagen; Valencia, CA), passed through a 22G needle five times, and then passed through a 27G needle five times. RNA was collected and isolated using the Micro-RNeasy Extraction kit (Qiagen), and cDNA was synthesized using the Qiagen FirstStrand Kit according to manufacturer’s instructions. qRT-PCR was performed using mouse primers for Mmp-1a, Mmp-2, Mmp-8, Mmp-9, Mmp-13, Mmp-14, and Gapdh (all Qiagen) with SsoAdvanced Universal SYBR Green Supermix (Bio-Rad; Hercules, CA). PCR was run on a CFX real time PCR machine (Bio-Rad) for a total of 40 cycles and three samples were run in duplicate for each condition. Data is expressed as fold change, with ± 2-fold set as the threshold for significance.
Fleszar A.J., Walker A., Kreeger P.K, & Notbohm J. (2019). Substrate curvature induces fallopian tube epithelial cell invasion via cell-cell tension in a model of ovarian cortical inclusion cysts. Integrative biology : quantitative biosciences from nano to macro, 11(8), 342-352.
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4

Extracellular Matrix Gene Expression Analysis

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RNA was collected and isolated using the Micro-RNeasy Extraction kit (Qiagen; Valencia, CA), and cDNA was synthesized using the Qiagen FirstStrand Kit according to manufacturer’s instructions. cDNA was mixed with Qiagen Mastermix and assayed using the Extracellular Matrix and Adhesion Molecules RT2 Profiler PCR Array (Qiagen) in a CFX real time PCR machine (Bio-Rad; Hercules, CA) for a total of 40 cycles, using Qiagen’s Data Analysis Center for analysis. Data is expressed as fold change, with ± 2-fold set as the threshold for significance. qRT-PCR was performed using human primers for SELP, CCR1, and CCR5, and GAPDH (all Qiagen), with SsoAdvanced Universal SYBR Green Supermix (Bio-Rad). Three samples were run in duplicate from each condition.
Carroll M.J., Fogg K.C., Patel H.A., Krause H.B., Mancha A.S., Patankar M.S., Weisman P.S., Barroilhet L, & Kreeger P.K. (2018). Alternatively activated macrophages upregulate mesothelial expression of P-selectin to enhance adhesion of ovarian cancer cells. Cancer research, 78(13), 3560-3573.
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5

Quantifying MMP9 Gene Expression

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RNA was collected using the Micro-RNeasy Extraction kit (Qiagen) and cDNA was synthesized at 60 ng/20 μL using SuperScript III First-Strand Synthesis System (Invitrogen). qRT-PCR was performed on 6 ng of cDNA using primers for MMP9 and GAPDH (Qiagen) and SsoAdvanced Universal SYBR Green Supermix (Bio-Rad), with three samples run in duplicate from each condition.
Carroll M.J., Kapur A., Felder M., Patankar M.S, & Kreeger P.K. (2016). M2 macrophages induce ovarian cancer cell proliferation via a heparin binding epidermal growth factor/matrix metalloproteinase 9 intercellular feedback loop. Oncotarget, 7(52), 86608-86620.
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6

Transcriptome Analysis of PC-3 Cells

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RNA was isolated from PC-3 cells using RNeasy micro-extraction kit (Qiagen, Germany) and quantified, and the purity was determined by measuring absorbance at 280, 260, and 230 nm using an ND-1000 nanodrop spectrophotometer (LabTech International, UK). After reverse transcription, cDNA was subjected to microarray analysis using the Human Genome U133 Plus 2.0 array (Affymetrix, High Wycombe, UK). Gene expression levels were analyzed using Microarray Suites software. Sequences used in the design of this array were selected from the GenBankTM, dbEST, and RefSeq databases.
Smith G.A., Howell G.J., Phillips C., Muench S.P., Ponnambalam S, & Harrison M.A. (2016). Extracellular and Luminal pH Regulation by Vacuolar H+-ATPase Isoform Expression and Targeting to the Plasma Membrane and Endosomes. The Journal of Biological Chemistry, 291(16), 8500-8515.
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7

Quantitative RT-PCR Analysis of Gene Expression

Cited 1 time
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Cells were starved in 2% FBS-EBM2 with and without inhibitors for 16 hours. Total cellular RNA was extracted with a RNeasy Micro extraction kit (Qiagen). Reverse transcriptase reactions were performed using an iScript™cDNA synthesis kit (BioRad). qPCR was performed using iTaq Universal SYBR Green Supermix (BioRad). Amplification was carried out in an ABI 7500 (Applied Biosystems). A relative standard curve of each gene amplification was generated to determine the amplification efficiency, with greater than 90% considered acceptable. Gene expression was normalized to GNAQ 27 (link). ATP5B and HPRTI were used as housekeeping genes. Quantitative analysis was performed according to delta CT based Pfaffl’s method 28 (link). Primer sequences are shown in Table 2.
Huang L., Bichsel C., Norris A., Thorpe J., Pevsner J., Alexandrescu S., Pinto A., Zurakowski D., Kleiman R.J., Sahin M., Greene A.K, & Bischoff J. (2021). Endothelial GNAQ p.R183Q increases angiopoietin-2 and drives formation of enlarged blood vessels. Arteriosclerosis, thrombosis, and vascular biology, 42(1), e27-e43.
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8

Quantitative PCR Analysis of Gene Expression

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Total cellular RNA was extracted from cells with an RNeasy Micro Extraction Kit (QIAGEN). Reverse transcriptase reactions were performed using an iScript cDNA Synthesis Kit (Bio-Rad). qPCR was performed using Kapa SYBR FAST ABI Prism 2× qPCR Master Mix (Kapa BioSystems). Amplification was carried out in a StepOne Real-Time PCR System (Applied Biosystems). A relative standard curve for each gene amplification was generated to determine the amplification efficiency, with greater than 90% considered acceptable. Fold increases in gene expression were calculated according to the ΔΔCt method, with each amplification reaction performed in duplicate or triplicate (69 (link), 70 (link)). Gene expression was normalized to the PBS treatment. ATP5B was used as housekeeping gene expression reference. Primer sequences are shown in Table 1.
Seebauer C.T., Graus M.S., Lan H.u.a.n.g., McCann A., Wylie-Sears J., Fontaine F., Karnezis T., Zurakowski D., Staffa S.J., Meunier F., Mulliken J.B., Bischoff J, & Francois M. (2022). Non–beta blocker enantiomers of propranolol and atenolol inhibit vasculogenesis in infantile hemangioma. The Journal of Clinical Investigation, 132(3), e151109.
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9

Quantitative Transcriptional Analysis of FACS-isolated Cells

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Total RNA was extracted according to the RNeasy microextraction kit (Qiagen, Hilden, Germany, #74004) from the FACS-isolated phenotypes. Extracted RNA was reverse-transcribed to generate first-strand cDNA (QuantiTect Reverse Transcription Kit, Qiagen, Hilden, Germany) for use in qPCR. qPCR was performed with Dnase-treated RNA using the Power SYBR Green Master Mix (Thermofisher Scientific, Waltham, MA, USA) on a real-time PCR system (Applied Biosystems, Waltham, MA, USA). qPCR primers were designed using PrimER-Blast software (http://www.ncbi.nlm.nih.gov/tools/primer-blast) and were based on previous studies. Supplementary Table S2 lists all primers used for this study.
Vikram R., Chou W.C., Hung S.C, & Shen C.Y. (2020). Tumorigenic and Metastatic Role of CD44−/low/CD24−/low Cells in Luminal Breast Cancer. Cancers, 12(5), 1239.
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10

Quantitative Gene Expression Analysis

Cited 1 time
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Total RNA was isolated using an RNeasy Micro extraction kit (Qiagen, Courtaboeuf, France), according to the manufacturer’s protocol. An on-column DNase I digestion was performed to avoid genomic DNA amplification. RNA level and quality were checked using the Nanodrop technology. A total of 500 ng of RNA was used for reverse transcription using the Superscript III reverse transcription kit (Invitrogen). Q-PCR analysis was performed using a QuantStudio 12 K Flex real-time PCR system (Applied biosystem) and TaqMan gene expression Master Mix (Roche), respectively, following the manufacturers’ instructions. Quantification of gene expression was based on the DeltaCt Method and normalized to 18 S expression. PCR primers for 18 S, MITF, TYRP1, PMEL17 lamin A, lamin C and progerin were previously described14 (link),25 (link),26 (link).
Lo Cicero A., Saidani M., Allouche J., Egesipe A.L., Hoch L., Bruge C., Sigaudy S., De Sandre-Giovannoli A., Levy N., Baldeschi C, & Nissan X. (2018). Pathological modelling of pigmentation disorders associated with Hutchinson-Gilford Progeria Syndrome (HGPS) revealed an impaired melanogenesis pathway in iPS-derived melanocytes. Scientific Reports, 8, 9112.
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