rneasy plus mini kit  (Qiagen)

 
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    Name:
    RNeasy Plus Mini Kit
    Description:
    For phenol free total RNA extraction from cells tissues with removal of genomic DNA contamination Kit contents Qiagen RNeasy Plus Mini Kit 50 preps 30mg Sample 30 to 50L Elution Volume Tissue Cells Sample Total RNA Purification Silica Technology Spin Column Format 25 min Time Run Ideal for PCR Northern Dot and Slot Blotting Array Analysis Sequencing For Purification of Up to 100g Total RNA from Cells tissues Using gDNA Eliminator Columns Includes RNeasy Mini Spin Columns gDNA Eliminator Spin Columns Collection Tubes RNase free Water and Buffers Benefits Efficient gDNA removal with unique gDNA Eliminator columns no need for DNase High quality total RNA in minutes using fast and simple extraction protocols Phenol free RNA isolation Ideal for sensitive applications such as real time RT PCR and RNA Seq
    Catalog Number:
    74134
    Price:
    360
    Category:
    RNeasy Plus Micro and Mini Kits
    Buy from Supplier


    Structured Review

    Qiagen rneasy plus mini kit
    RNeasy Plus Mini Kit
    For phenol free total RNA extraction from cells tissues with removal of genomic DNA contamination Kit contents Qiagen RNeasy Plus Mini Kit 50 preps 30mg Sample 30 to 50L Elution Volume Tissue Cells Sample Total RNA Purification Silica Technology Spin Column Format 25 min Time Run Ideal for PCR Northern Dot and Slot Blotting Array Analysis Sequencing For Purification of Up to 100g Total RNA from Cells tissues Using gDNA Eliminator Columns Includes RNeasy Mini Spin Columns gDNA Eliminator Spin Columns Collection Tubes RNase free Water and Buffers Benefits Efficient gDNA removal with unique gDNA Eliminator columns no need for DNase High quality total RNA in minutes using fast and simple extraction protocols Phenol free RNA isolation Ideal for sensitive applications such as real time RT PCR and RNA Seq
    https://www.bioz.com/result/rneasy plus mini kit/product/Qiagen
    Average 99 stars, based on 1224 article reviews
    Price from $9.99 to $1999.99
    rneasy plus mini kit - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment"

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0214609

    Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.
    Figure Legend Snippet: Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.

    Techniques Used: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Isolation, Molecular Weight, Marker

    Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p
    Figure Legend Snippet: Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Techniques Used: Real-time Polymerase Chain Reaction, Isolation

    2) Product Images from "Ectromelia virus accumulates less double-stranded RNA compared to vaccinia virus in BS-C-1 cells"

    Article Title: Ectromelia virus accumulates less double-stranded RNA compared to vaccinia virus in BS-C-1 cells

    Journal: Virology

    doi: 10.1016/j.virol.2017.06.010

    The accumulation of dsRNA greatly varies after infection with different orthopoxviruses (A) BS-C-1 cells were infected (MOI=10) with ECTV, VACV, or CPXV for 24 hrs. prior to staining for dsRNA (red) and cell nuclei/virus factories (DAPI; blue). Images were merged using ImageJ software, are at 400× total magnification, and representative of three independent trials. AraC was added at the time of infection where indicated. (B) BS-C-1 cells were infected (MOI=10) with either ECTV or VACV for 24 hrs. prior to staining intracellularly for dsRNA and analyzed using flow cytometry. Positive gates were drawn based upon the mock condition. The depicted data are representative of four independent trials. (C) Flow cytometry was used to measure dsRNA levels within infected cells as above. The data are shown in histogram format and representative of four independent trials. (D) BS-C-1 cells were infected (MOI=10) with ECTV or VACV. RNA was isolated at 24 hrs. post-infection using the RNeasy Plus Mini Kit (Qiagen) and 5 µg RNA was spotted onto a membrane. dsRNA was detected using the J2 antibody and beta-actin mRNA was detected using a biotin-labeled antisense RNA as described in the methods.
    Figure Legend Snippet: The accumulation of dsRNA greatly varies after infection with different orthopoxviruses (A) BS-C-1 cells were infected (MOI=10) with ECTV, VACV, or CPXV for 24 hrs. prior to staining for dsRNA (red) and cell nuclei/virus factories (DAPI; blue). Images were merged using ImageJ software, are at 400× total magnification, and representative of three independent trials. AraC was added at the time of infection where indicated. (B) BS-C-1 cells were infected (MOI=10) with either ECTV or VACV for 24 hrs. prior to staining intracellularly for dsRNA and analyzed using flow cytometry. Positive gates were drawn based upon the mock condition. The depicted data are representative of four independent trials. (C) Flow cytometry was used to measure dsRNA levels within infected cells as above. The data are shown in histogram format and representative of four independent trials. (D) BS-C-1 cells were infected (MOI=10) with ECTV or VACV. RNA was isolated at 24 hrs. post-infection using the RNeasy Plus Mini Kit (Qiagen) and 5 µg RNA was spotted onto a membrane. dsRNA was detected using the J2 antibody and beta-actin mRNA was detected using a biotin-labeled antisense RNA as described in the methods.

    Techniques Used: Infection, Staining, Software, Flow Cytometry, Cytometry, Isolation, Labeling

    3) Product Images from "Activation of Melanocortin Receptors MC1 and MC5 Attenuates Retinal Damage in Experimental Diabetic Retinopathy"

    Article Title: Activation of Melanocortin Receptors MC1 and MC5 Attenuates Retinal Damage in Experimental Diabetic Retinopathy

    Journal: Mediators of Inflammation

    doi: 10.1155/2016/7368389

    Real Time PCR for the expression of melanocortin receptor subtypes within the retina. (a) Representative traces of the RT-PCR and (b) relative 2 −ΔΔCt gene expressions for MC 1 , MC 3 , and MC 5 receptors assayed after 16-week follow-up in nondiabetic mice, and diabetic mice with retinopathy. Total RNA was extracted using RNeasy Plus Mini Kit and commercially available primer for amplification of mouse MC 1 , MC 3 , and MC 5 receptors. Negative controls were either RT without enzyme or PCR without cDNA template.
    Figure Legend Snippet: Real Time PCR for the expression of melanocortin receptor subtypes within the retina. (a) Representative traces of the RT-PCR and (b) relative 2 −ΔΔCt gene expressions for MC 1 , MC 3 , and MC 5 receptors assayed after 16-week follow-up in nondiabetic mice, and diabetic mice with retinopathy. Total RNA was extracted using RNeasy Plus Mini Kit and commercially available primer for amplification of mouse MC 1 , MC 3 , and MC 5 receptors. Negative controls were either RT without enzyme or PCR without cDNA template.

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Reverse Transcription Polymerase Chain Reaction, Mouse Assay, Amplification, Polymerase Chain Reaction

    4) Product Images from "Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment"

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0214609

    Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p
    Figure Legend Snippet: Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Techniques Used: Real-time Polymerase Chain Reaction, Isolation

    5) Product Images from "Ectromelia virus accumulates less double-stranded RNA compared to vaccinia virus in BS-C-1 cells"

    Article Title: Ectromelia virus accumulates less double-stranded RNA compared to vaccinia virus in BS-C-1 cells

    Journal: Virology

    doi: 10.1016/j.virol.2017.06.010

    The accumulation of dsRNA greatly varies after infection with different orthopoxviruses (A) BS-C-1 cells were infected (MOI=10) with ECTV, VACV, or CPXV for 24 hrs. prior to staining for dsRNA (red) and cell nuclei/virus factories (DAPI; blue). Images were merged using ImageJ software, are at 400× total magnification, and representative of three independent trials. AraC was added at the time of infection where indicated. (B) BS-C-1 cells were infected (MOI=10) with either ECTV or VACV for 24 hrs. prior to staining intracellularly for dsRNA and analyzed using flow cytometry. Positive gates were drawn based upon the mock condition. The depicted data are representative of four independent trials. (C) Flow cytometry was used to measure dsRNA levels within infected cells as above. The data are shown in histogram format and representative of four independent trials. (D) BS-C-1 cells were infected (MOI=10) with ECTV or VACV. RNA was isolated at 24 hrs. post-infection using the RNeasy Plus Mini Kit (Qiagen) and 5 µg RNA was spotted onto a membrane. dsRNA was detected using the J2 antibody and beta-actin mRNA was detected using a biotin-labeled antisense RNA as described in the methods.
    Figure Legend Snippet: The accumulation of dsRNA greatly varies after infection with different orthopoxviruses (A) BS-C-1 cells were infected (MOI=10) with ECTV, VACV, or CPXV for 24 hrs. prior to staining for dsRNA (red) and cell nuclei/virus factories (DAPI; blue). Images were merged using ImageJ software, are at 400× total magnification, and representative of three independent trials. AraC was added at the time of infection where indicated. (B) BS-C-1 cells were infected (MOI=10) with either ECTV or VACV for 24 hrs. prior to staining intracellularly for dsRNA and analyzed using flow cytometry. Positive gates were drawn based upon the mock condition. The depicted data are representative of four independent trials. (C) Flow cytometry was used to measure dsRNA levels within infected cells as above. The data are shown in histogram format and representative of four independent trials. (D) BS-C-1 cells were infected (MOI=10) with ECTV or VACV. RNA was isolated at 24 hrs. post-infection using the RNeasy Plus Mini Kit (Qiagen) and 5 µg RNA was spotted onto a membrane. dsRNA was detected using the J2 antibody and beta-actin mRNA was detected using a biotin-labeled antisense RNA as described in the methods.

    Techniques Used: Infection, Staining, Software, Flow Cytometry, Cytometry, Isolation, Labeling

    6) Product Images from "Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment"

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0214609

    Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p
    Figure Legend Snippet: Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Techniques Used: Real-time Polymerase Chain Reaction, Isolation

    7) Product Images from "Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment"

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0214609

    Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.
    Figure Legend Snippet: Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.

    Techniques Used: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Isolation, Molecular Weight, Marker

    Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p
    Figure Legend Snippet: Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Techniques Used: Real-time Polymerase Chain Reaction, Isolation

    8) Product Images from "Mutated NPM1 in combination with overexpression of Meis1 or Hoxa9 is not sufficient to induce acute myeloid leukemia"

    Article Title: Mutated NPM1 in combination with overexpression of Meis1 or Hoxa9 is not sufficient to induce acute myeloid leukemia

    Journal: Experimental Hematology & Oncology

    doi: 10.1186/s40164-016-0053-2

    Transduction of murine bone marrow cells with NPMc + , Meis1 and Hoxa9. a The table shows the combination of genes murine bone marrow cells were transduced with. b – d RNA was extracted from transduced cells using the RNeasy Plus Mini kit and analyzed by qPCR for expression of human NPM1 ( b ), human MEIS1 ( c ) and murine Hoxa9 ( d ). For all qPCR analysis n = 3. The bars indicate mean ± SD. Statistical calculations were performed using Student´s unpaired t test. *p
    Figure Legend Snippet: Transduction of murine bone marrow cells with NPMc + , Meis1 and Hoxa9. a The table shows the combination of genes murine bone marrow cells were transduced with. b – d RNA was extracted from transduced cells using the RNeasy Plus Mini kit and analyzed by qPCR for expression of human NPM1 ( b ), human MEIS1 ( c ) and murine Hoxa9 ( d ). For all qPCR analysis n = 3. The bars indicate mean ± SD. Statistical calculations were performed using Student´s unpaired t test. *p

    Techniques Used: Transduction, Real-time Polymerase Chain Reaction, Expressing

    9) Product Images from "Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment"

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0214609

    Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.
    Figure Legend Snippet: Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.

    Techniques Used: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Isolation, Molecular Weight, Marker

    Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p
    Figure Legend Snippet: Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Techniques Used: Real-time Polymerase Chain Reaction, Isolation

    10) Product Images from "Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment"

    Article Title: Elimination of bacterial DNA during RNA isolation from sputum: Bashing bead vortexing is preferable over prolonged DNase treatment

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0214609

    Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.
    Figure Legend Snippet: Representative gel electrophoresis of PCR reactions using universal bacterial primers and sputum RNA isolated with the RNeasy Plus Mini kit. Sputum cells were either subjected or not to bead vortexing (bashing or glass beads) prior to RNA isolation. Bands representing contaminating bacterial DNA are indicated. M: molecular weight marker.

    Techniques Used: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Isolation, Molecular Weight, Marker

    Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p
    Figure Legend Snippet: Cycle threshold (Ct) values for real-time PCR assays with different base pair (bp) long amplicons of glyceraldehyde 3-phophate dehydrogenase on RNA isolated from sputum samples. Samples were subjected either to repeated digestion process (up to 6 times) with the Turbo DNA-free kit or bead vortexing prior to RNA isolation using the RNeasy Plus Mini kit equipped with the gDNA eliminator spin column (n = 6 for each group). Error bars indicate SEM. # p

    Techniques Used: Real-time Polymerase Chain Reaction, Isolation

    Related Articles

    Transfection:

    Article Title: The Tumor Suppressive Role of eIF3f and Its Function in Translation Inhibition and rRNA Degradation
    Article Snippet: .. Cells were harvested at 4, 8 and 16 h after transfection and total RNA was isolated using RNeasy kit (QIAGEN). rRNAs (2.0 µg) were separated on an agarose gel and visualized by UV light (B). ..

    Agarose Gel Electrophoresis:

    Article Title: The Tumor Suppressive Role of eIF3f and Its Function in Translation Inhibition and rRNA Degradation
    Article Snippet: .. Cells were harvested at 4, 8 and 16 h after transfection and total RNA was isolated using RNeasy kit (QIAGEN). rRNAs (2.0 µg) were separated on an agarose gel and visualized by UV light (B). ..

    Isolation:

    Article Title: Research Resource: A Genome-Wide Study Identifies Potential New Target Genes for POU1F1
    Article Snippet: .. Total RNA from cells infected with the lentiviral vectors was isolated by using the RNeasy Kit (Qiagen, Hilden, Germany), including on-column DNase I digestion according to the manufacturer's recommendations. .. The quality of isolated RNA was controlled by the Lab-on-Chip-System Bioanalyser 2100 (Agilent Technologies, Inc., Palo Alto, CA).

    Article Title: The Tumor Suppressive Role of eIF3f and Its Function in Translation Inhibition and rRNA Degradation
    Article Snippet: .. Cells were harvested at 4, 8 and 16 h after transfection and total RNA was isolated using RNeasy kit (QIAGEN). rRNAs (2.0 µg) were separated on an agarose gel and visualized by UV light (B). ..

    Article Title: The embryonic type of SPP1 transcriptional regulation is re-activated in glioblastoma
    Article Snippet: .. Real- time PCR Total RNA was isolated from glioma cells using Qiagen RNeasy kit, 1 μg was used as a template. .. Amplifications were performed in duplicates in 20 μl containing 2× SYBR PCR MasterMix (Applied Biosystems) and a set of primers (human SPP1, NANOG and POU5F1_1_SG/OCT4 primers were from QuantiTect Primer Assays: QT01008798, QT01844808 and QT00210840, respectively, Qiagen).

    Article Title: Mechanisms by which Porphyromonas gingivalis evades innate immunity
    Article Snippet: .. Quantitative RT-PCR BMDCs were cultured in a 24-well plate for 6 h with 108 bacteria/well and RNA was isolated from the cells by RNeasy kit (Qiagen). .. RNA was treated to remove DNA contamination (Invitrogen) and the cDNA was synthesized using random primers, and reverse transcribed using Superscript III (Invitrogen).

    Infection:

    Article Title: Research Resource: A Genome-Wide Study Identifies Potential New Target Genes for POU1F1
    Article Snippet: .. Total RNA from cells infected with the lentiviral vectors was isolated by using the RNeasy Kit (Qiagen, Hilden, Germany), including on-column DNase I digestion according to the manufacturer's recommendations. .. The quality of isolated RNA was controlled by the Lab-on-Chip-System Bioanalyser 2100 (Agilent Technologies, Inc., Palo Alto, CA).

    Purification:

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels
    Article Snippet: .. The protocol developed by Wang et al. uses a combination of mechanical disruption, extraction with CTAB buffer and purification with the RNeasy® kit. .. To test the efficacy of this approach with our specific material and cell source, the hydrogels were either (1) cryo-pulverized with a mortar and pestle into a fine powder or (2) finely minced with sharp surgical scissors and then disrupted using an ultrasonic dismembrator (Fisher Scientific Model 100) with three 5-s bursts at a setting of 4 with intervals of cooling on ice in 600 μL of CTAB buffer [2% CTAB, 2% poly(vinylpyrrolidone) (PVP), 1.4 M NaCl, 100 mM Tris-HCl, 20 mM EDTA, and 1% beta-mercaptoethanol in diethylpyrocarbonate (DEPC)-treated water] prewarmed to 65°C.

    Real-time Polymerase Chain Reaction:

    Article Title: The embryonic type of SPP1 transcriptional regulation is re-activated in glioblastoma
    Article Snippet: .. Real- time PCR Total RNA was isolated from glioma cells using Qiagen RNeasy kit, 1 μg was used as a template. .. Amplifications were performed in duplicates in 20 μl containing 2× SYBR PCR MasterMix (Applied Biosystems) and a set of primers (human SPP1, NANOG and POU5F1_1_SG/OCT4 primers were from QuantiTect Primer Assays: QT01008798, QT01844808 and QT00210840, respectively, Qiagen).

    Cell Culture:

    Article Title: Mechanisms by which Porphyromonas gingivalis evades innate immunity
    Article Snippet: .. Quantitative RT-PCR BMDCs were cultured in a 24-well plate for 6 h with 108 bacteria/well and RNA was isolated from the cells by RNeasy kit (Qiagen). .. RNA was treated to remove DNA contamination (Invitrogen) and the cDNA was synthesized using random primers, and reverse transcribed using Superscript III (Invitrogen).

    Quantitative RT-PCR:

    Article Title: Mechanisms by which Porphyromonas gingivalis evades innate immunity
    Article Snippet: .. Quantitative RT-PCR BMDCs were cultured in a 24-well plate for 6 h with 108 bacteria/well and RNA was isolated from the cells by RNeasy kit (Qiagen). .. RNA was treated to remove DNA contamination (Invitrogen) and the cDNA was synthesized using random primers, and reverse transcribed using Superscript III (Invitrogen).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Long non-coding RNA MUC5B-AS1 promotes metastasis through mutually regulating MUC5B expression in lung adenocarcinoma
    Article Snippet: .. We then cleaned up RNA using RNeasy kits (QIAGEN, Madison, WI, USA) and carried out reverse transcription-polymerase chain reaction (RT-PCR) using the primers to detect the OL and non-OL regions of the MUC5B mRNA. ..

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  • 99
    Qiagen rneasy kit
    Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total <t>RNA</t> was harvested from the peripheral blood mononuclear cells using an <t>RNeasy</t> kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.
    Rneasy Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2846 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy kit/product/Qiagen
    Average 99 stars, based on 2846 article reviews
    Price from $9.99 to $1999.99
    rneasy kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

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    Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total RNA was harvested from the peripheral blood mononuclear cells using an RNeasy kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Therapeutic effects of calcium dobesilate on diabetic nephropathy mediated through reduction of expression of PAI-1

    doi: 10.3892/etm.2012.755

    Figure Lengend Snippet: Quantitative RT-PCR analysis of PAI-1 mRNA levels in patients. The total RNA was harvested from the peripheral blood mononuclear cells using an RNeasy kit according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system according to the manufacturer’s instructions. The expression of PAI-1 mRNA was quantified by quantitative PCR using an ABI Prism Sequence Detection System. Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. The levels (mean value) of PAI-1 transcripts in patients were calculated. RT-PCR, reverse-transcription-PCR, PAI-1, plasminogen activator inhibitor-1.

    Article Snippet: The total RNA was harvested from peripheral blood mononuclear cells using the RNeasy kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.

    Techniques: Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction, Sequencing, Amplification, Polymerase Chain Reaction

    2′- O methylation of internal adenosine. (A) Incorporation of 3 H-methyl into polyA. Homopolymer RNAs (1 µg) were incubated with 2 µg of DENV-4 MTase in the presence of [ 3 H-methyl]-SAM. After the methylation reaction, the un-incorporated [ 3 H-methyl]-SAM was removed by RNeasy kit. The amount of 3 H-methyl incorporation was measured by a MicroBeta counting. (B) SPA-based methylation analysis of oligo (A) 12 , (Am) 12 , and (m 6 ,m 6 A) 12 . All three RNA oligos were 3′-end biotinylated to facilitate SPA analysis. Am indicates that the 2′-OH of adenosine is methylated. m 6 ,m 6 A indicates that the amino N 6 position of adenosine is double methylated. (C) SPA-based methylation analysis of DENV-1 RNA. pppA-RNAs, representing the 5′ 211 nt of DENV-1 genome, were in vitro transcribed using biotinylated-CTP plus unmodified ATP, 2′- O -methyladenosine triphosphate (AmTP), or N 6 methyl adenosine triphosphate (m 6 ATP). The transcription reactions generated pppA-RNA, ppp(Am)-RNA, and ppp(m 6 A)RNA, respectively. The RNAs were then subjected to SPA-based internal methylation analysis. Average results and standard deviations from three independent experiments are presented.

    Journal: PLoS Pathogens

    Article Title: 2?-O Methylation of Internal Adenosine by Flavivirus NS5 Methyltransferase

    doi: 10.1371/journal.ppat.1002642

    Figure Lengend Snippet: 2′- O methylation of internal adenosine. (A) Incorporation of 3 H-methyl into polyA. Homopolymer RNAs (1 µg) were incubated with 2 µg of DENV-4 MTase in the presence of [ 3 H-methyl]-SAM. After the methylation reaction, the un-incorporated [ 3 H-methyl]-SAM was removed by RNeasy kit. The amount of 3 H-methyl incorporation was measured by a MicroBeta counting. (B) SPA-based methylation analysis of oligo (A) 12 , (Am) 12 , and (m 6 ,m 6 A) 12 . All three RNA oligos were 3′-end biotinylated to facilitate SPA analysis. Am indicates that the 2′-OH of adenosine is methylated. m 6 ,m 6 A indicates that the amino N 6 position of adenosine is double methylated. (C) SPA-based methylation analysis of DENV-1 RNA. pppA-RNAs, representing the 5′ 211 nt of DENV-1 genome, were in vitro transcribed using biotinylated-CTP plus unmodified ATP, 2′- O -methyladenosine triphosphate (AmTP), or N 6 methyl adenosine triphosphate (m 6 ATP). The transcription reactions generated pppA-RNA, ppp(Am)-RNA, and ppp(m 6 A)RNA, respectively. The RNAs were then subjected to SPA-based internal methylation analysis. Average results and standard deviations from three independent experiments are presented.

    Article Snippet: For each time point, total cellular RNA was extracted using RNeasy kit (Qiagen).

    Techniques: Methylation, Incubation, In Vitro, Generated

    Inflammatory cytokine production from DCs is extinguished in the presence of P . gingivalis . ( A ) DCs from B10.A-Rag2 -/- mice were either unstimulated (medium) or stimulated on the sixth day with 5x10 7 bacteria in a 24-well plate for 16h at 37°C. Cells were harvested, stained with directly labeled monoclonal antibodies and analyzed by FACS analysis. Cells were gated for the CD11c positive population and analyzed for the costimulatory molecules B7.1, B7.2 and CD40, and the antigen-presenting molecule, MHC class II. Numbers indicate the percentage of the population in the gate or quadrant ( B ) Same as (A) Culture CSN were analyzed for the presence of various cytokines using Searchlight protein arrays. ( C ) Same as (B) except the co-culture period was 6h, at which time we isolated total RNA using the RNeasy kit and analyzed it by Real-time RT-PCR. The data are expressed as the mean ± SD of three independent experiments.

    Journal: PLoS ONE

    Article Title: Mechanisms by which Porphyromonas gingivalis evades innate immunity

    doi: 10.1371/journal.pone.0182164

    Figure Lengend Snippet: Inflammatory cytokine production from DCs is extinguished in the presence of P . gingivalis . ( A ) DCs from B10.A-Rag2 -/- mice were either unstimulated (medium) or stimulated on the sixth day with 5x10 7 bacteria in a 24-well plate for 16h at 37°C. Cells were harvested, stained with directly labeled monoclonal antibodies and analyzed by FACS analysis. Cells were gated for the CD11c positive population and analyzed for the costimulatory molecules B7.1, B7.2 and CD40, and the antigen-presenting molecule, MHC class II. Numbers indicate the percentage of the population in the gate or quadrant ( B ) Same as (A) Culture CSN were analyzed for the presence of various cytokines using Searchlight protein arrays. ( C ) Same as (B) except the co-culture period was 6h, at which time we isolated total RNA using the RNeasy kit and analyzed it by Real-time RT-PCR. The data are expressed as the mean ± SD of three independent experiments.

    Article Snippet: Quantitative RT-PCR BMDCs were cultured in a 24-well plate for 6 h with 108 bacteria/well and RNA was isolated from the cells by RNeasy kit (Qiagen).

    Techniques: Mouse Assay, Staining, Labeling, FACS, Co-Culture Assay, Isolation, Quantitative RT-PCR

    Representative end-point RT-PCR gene expression results for (a) 18S and (b) TfR from RNA isolated using the freeze grind+CTAB+RNeasy ® (FCR) method, the mince+CTAB+RNeasy ® (MCR) method, and the lysozyme+CTAB+RNeasy ® (LCR) method. Genomic contamination was detected following agarose gel electrophoresis in all minus-RT controls. In addition, as shown in the TfR results, where the primers were designed to span an intron–exon boundary, two products were formed during the PCR, corresponding to a genomic product size of 270 bp and an mRNA product size of 62 bp.

    Journal: Tissue Engineering. Part C, Methods

    Article Title: Techniques for the Isolation of High-Quality RNA from Cells Encapsulated in Chitosan Hydrogels

    doi: 10.1089/ten.tec.2012.0693

    Figure Lengend Snippet: Representative end-point RT-PCR gene expression results for (a) 18S and (b) TfR from RNA isolated using the freeze grind+CTAB+RNeasy ® (FCR) method, the mince+CTAB+RNeasy ® (MCR) method, and the lysozyme+CTAB+RNeasy ® (LCR) method. Genomic contamination was detected following agarose gel electrophoresis in all minus-RT controls. In addition, as shown in the TfR results, where the primers were designed to span an intron–exon boundary, two products were formed during the PCR, corresponding to a genomic product size of 270 bp and an mRNA product size of 62 bp.

    Article Snippet: The protocol developed by Wang et al. uses a combination of mechanical disruption, extraction with CTAB buffer and purification with the RNeasy® kit.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Isolation, Agarose Gel Electrophoresis, Polymerase Chain Reaction